PQDx 0156 053 00 WHO PQDx PR November/2015, … 0156‐053‐00 WHO PQDx PR November/2015, version...

PQDx 0156‐053‐00 WHO PQDx PR November/2015, version 2.0

of 40

WHO Prequalification of Diagnostics Programme

PUBLIC REPORT

Product: Aquios CL flow cytometer Number: PQDx 0156‐053‐00

Abstract

Aquios CL flow cytometer with the product codes listed below manufactured by Beckman Coulter Life Sciences, CE marked regulatory version, was accepted for the WHO list of prequalified in vitro diagnostics and was listed on 9 November 2015. Intended use: Aquios CL cytometer together with AQUIOS Tetra‐1 Panel (CD45‐FITC/CD4‐RD1/CD8‐ECD/CD3‐PC5) and Aquios Immuno‐Trol/Immuno‐Trol low controls are intended for use with in‐vitro diagnostic flow cytometric applications (four fluorescent detection channels using a blue (488 nm) laser, two light scatter detection channels and electronic volume (EV). The system is used for immunologic assessment of patients having, or suspected of having, immune deficiency. These reagents provide identification and enumeration of the following lymphocyte subset populations:

total CD3+, CD3+CD4+,CD3+CD8+, CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts;

CD45+ absolute count; and

CD45+ Low SS (lymphocytes) percentage and absolute count.

An additional reagent is available, however, it was not reviewed as part of this WHO prequalification assessment ‐ AQUIOS Tetra‐2+ Panel monoclonal antibody reagents that identified and enumerates the following lymphocyte subset populations: Total CD3+, CD3‐CD19+, CD3‐CD56+ and/or CD16+ lymphocyte percentages and absolute counts; CD45+ absolute count; and CD45+ Low SS (lymphocytes) percentage and absolute count.

Assay principle: The Aquios system can be operated using up to 40 samples at start‐up with the ability to add 5‐tube cassettes as testing proceeds. The tube(s) are loaded into the instrument cassette which is further loaded on the system including the Tetra‐1 Panel. Whole blood is incubated with AQUIOS Tetra‐1 Panel monoclonal antibody reagents which contains a combination of four murine monoclonal antibodies, (CD45‐FITC/CD4‐RD1/CD8‐ECD/CD3‐PC5). Each monoclonal antibody is conjugated to a specific fluorochrome and specific for different cell surface antigens. Red blood cells are lysed with the AQUIOS Lysing Reagent


Kit. The remaining white blood cells are analyzed by flow cytometry using lymphocyte gates. The AQUIOS System Software with AQUIOS Tetra‐1 Panel monoclonal antibody reagents provides automated analysis of lymphocyte subpopulations including total CD3+, CD3+CD4+,CD3+CD8+, CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts. In addition, CD45+ absolute count CD45+ Low SS (lymphocytes) percentage and absolute count. Limitation: Users may receive system notifications of “potential specimen or gating issue” that may require intervention and technical expertise to review the results before release. Notifications alert the operator that there may be a potential anomaly with the specimen result produced. Therefore it should be reviewed especially for the presentation of the cellular populations and the gating accomplished by the instrument, to verify that there are no additional changes needed, prior to releasing the result. In this prequalification assessment, the frequency of such notifications which required user intervention was up to 30% of all specimens tested at one of two sites. The system instrument:

Product name Product code Description

AQUIOS CL flow cytometer

B30166 Flow cytometry instrument

AQUIOS CL with UPS for 110v, AQUIOS Workstation and Software, Supply Cart

B39101 (1) flow cytometry instrument; (2) uninterruptible power supply for 110v (B37943); (3) all‐in‐one computer with touch‐screen monitor, wireless mouse, wireless keyboard; (4) vacuum pump module, vacuum trap bottle, sheath filter and sheath pump, and holds the AQUIOS Sheath Solution Cubitainer, and waste container

AQUIOS CL with UPS for 220v, AQUIOS Workstation and software, Supply Cart

B39102 (1) flow cytometry instrument; (2) uninterruptible power supply for 220v (B37954), cable assembly (6009252), interconnect cables (B43588) (3) all‐in‐one computer with touch‐screen monitor, wireless mouse, wireless keyboard; (4) vacuum pump module, vacuum trap bottle, sheath filter and sheath pump, and holds the AQUIOS Sheath Solution Cubitainer, and waste container

of 40


The system consumables:

Product name Product code

Quantity Description

AQUIOS Sheath Solution

B25697 1 x 10L A non‐fluorescent, azide‐free balanced electrolyte solution for use on AQUIOS CL flow cytometer with light scatter and fluorescent applications.

AQUIOS Cleaning Agent

B25698 1 x 500ml This azide‐free, formaldehyde‐free, biodegradable cleaner contains a proteolytic enzyme that aids in the removal of protein build‐up in the fluidics system and flow cell of a flow cytometer.

AQUIOS Sodium Hypochlorite Solution

B23536 4 X 50ml A cleaning agent that is used during cleaning cycles to counteract protein build‐up that occurs in tubing and the flow cell.

AQUIOS Lysing Reagent Kit

B23538 1 x 38ml 1 x 15ml (100 tests)

A cyanide‐free lytic reagent that lyses red blood cells to allow enumeration of white blood cells.

AQUIOS Tetra‐1 Panel B23533 1 x 0.9ml (50 tests)

Reagent panel that employs monoclonal antibodies: CD45‐FITC/CD4‐RD1/CD8‐ECD/CD3‐PC5.

AQUIOS Tetra‐2+ Panel B23534 1 x 0.9ml (50 tests)

Reagent panel that employs monoclonal antibodies: CD45‐FITC/CD56‐RD1+CD16‐RD1/CD19‐ ECD/CD3‐PC5. Note: this reagent was not reviewed as part of this prequalification assessment

AQUIOS IMMUNO‐TROL Cells

B23535 2 X 3mL An integrated control that enables monitoring of system performance for all directly measured and calculated parameters.

AQUIOS IMMUNO‐TROL LOW Cells

B25700 2 X 3mL An integrated control that enables monitoring of system performance for all directly measured and calculated parameters.

AQUIOS Deep Well Plates

B23502 50 plates/ box

Reservoirs for specimen preparation.

of 40


Accessories used as part of the system:

Accessories part

Product code

Quantity Provided/not provided with the instrument

Type 1 Cassette (13 x 75mm) B25218 2 Provided (additional

available for order)





Type 4 Cassette (10.25 x 50mm) B25221 2 Provided (additional


Type 5 Cassette (Immuno‐Trol) B25318 2 Provided (additional


Type 6 Cassette (Sarstedt 13 x 75mm) B25918 2 Provided (additional


Type 7 Cassette (Sarstedt 13 x 90mm) B52994 1 Not provided



Type 10 Cassette (Sarstedt 13 x 65mm)

B53808 1 Not provided

Color Laser Printer B31892 1 Not provided

4mL Round Bottom Test Tubes B42830 25 Provided

Gravimetric Bottle B42831 2 Provided

40 Test Tube Rack 178696 1 Provided

of 40


Storage conditions:

Product name Product code Temperature °C

AQUIOS CL Flow Cytometer B30166 N/A

AQUIOS Sheath Solution B25697 18 ‐ 26°C

AQUIOS Cleaning Agent B25698 18 ‐ 26°C


B23536 18 ‐ 26°C

AQUIOS Lysing Reagent Kit B23538 18 ‐ 26°C

AQUIOS Tetra‐1 Panel B23533 2 ‐ 8°C

AQUIOS Tetra‐2+ Panel B23534 2 ‐ 8°C

AQUIOS IMMUNO‐TROL Cells B23535 2 ‐ 8°C

AQUIOS IMMUNO‐TROL LOW Cells B25700 2 ‐ 8°C

AQUIOS Deep Well Plates B23502 N/A

Shelf‐life:

Product name Product code Shelf life

AQUIOS CL Flow Cytometer B30166 N/A

AQUIOS Sheath Solution B25697 18 months

AQUIOS Cleaning Agent B25698 12 months


B23536 12 months

AQUIOS Lysing Reagent Kit B23538 350 days

AQUIOS Tetra‐1 Panel B23533 12 months

AQUIOS Tetra‐2+ Panel B23534 12 months

AQUIOS IMMUNO‐TROL Cells B23535 270 days

AQUIOS IMMUNO‐TROL LOW Cells B25700 270 days

AQUIOS Deep Well Plates B23502 N/A

of 40


Summary of prequalification status for AQUIOS CL flow cytometer

Initial acceptance

Date Outcome

Status on PQ list 6 November 2015 listed

Dossier assessment 29 May 2015 MR

Inspection status 25 March 2015 MR

Laboratory evaluation 29 September 2015 MR

MR: Meets Requirements N/A: Not Applicable AQUIOS CL flow cytometer was accepted for the WHO list of prequalified in vitro diagnosticson the basis of data submitted and publicly available information. Background information Beckman Coulter Life Sciences submitted an application for prequalification of AQUIOS CL flow cytometer. Based on the established prioritization criteria, AQUIOS CL flow cytometer was given priority for prequalification. Product dossier assessment Beckman Coulter Life Sciences submitted a product dossier for AQUIOS CL flow cytometer as per the Instructions for compilation of a product dossier (PQDx_018 v1). The information submitted in the product dossier was reviewed by WHO staff and external experts (assessors) appointed by WHO in accordance with the internal report on the screening and assessment of a product dossier (PQDx_009 v2). Based on the product dossier screening and assessment findings, a recommendation was made to accept the product dossier for AQUIOS CL flow cytometer for prequalification. Manufacturing site inspection A comprehensive inspection was performed at the site of manufacture (11800 SW 147 Ave, Miami, FL, USA (instrument site) and 740 W 83 Street, Hialeah, FL, USA (reagent site) of AQUIOS CL flow cytometer in February 2015 as per the Information for manufacturers on prequalification inspection procedures for the sites of manufacture of diagnostics (PQDx_014 v1). The inspection found that the manufacturer had an acceptable quality management system and good manufacturing practices in place that ensured the consistent manufacture of a product of good quality. The manufacturer's responses to the nonconformities found at the time of the inspection were accepted 25 March 2015. Laboratory evaluation Aquios CL together with AQUIOS Tetra‐1 Panel (CD45‐FITC/CD4‐RD1/CD8‐ECD/CD3‐PC) and Aquios Immuno‐Trol and Aquios Immuno‐Trol low controls were evaluated in two WHO collaborating laboratories; namely Institute of Tropical Medicine (ITM), Antwerp, Belgium and Centres for Disease Control and Prevention, Atlanta, GA. USA (CDC) between

of 40


March and July 2015. The evaluation was conducted using the WHO evaluation protocol “Protocol for multicenter laboratory assessment of dedicated and point‐of‐care CD4+ T‐lymphocytes enumeration technologies” (SOP_PQDx_114) which was also approved by in‐country ethical review boards in Belgium and USA. Aquios CL together with AQUIOS Tetra‐1 Panel (CD45‐FITC/CD4‐RD1/CD8‐ECD/CD3‐PC) and Aquios Immuno‐Trol/Immuno‐Trol low controls are a bench‐top automated flow cytometric system intended for enumeration of CD3+, CD3+CD4+, CD3+CD8+, and CD3+CD4+/CD3+CD8+ (ratio only). 100 µl of well mixed venous whole blood and AQUIOSTM Tetra‐1 Panel reagents are required to perform the assays and the results are obtained after 15 minutes incubation. A total of 537 venous whole blood specimens were used study failure rate, reproducibility (intra‐laboratory variation and inter‐assay), carry over and agreement with the FACSCalibur™ as the reference method. Lastly, ease to use was assessed. The acceptance criteria for reproducibility studies was that the assay should have the a percentage coefficient of variation (%CV) less than 15% for CD4+ T counts less than or equal to 200/µL and less than 10% for CD4 counts more than 200 cells/µL, while the carry‐over constant (k) should be less than 2.0%. Consecutive routine venous whole blood specimens collected in EDTA vacutainer tubes with at least 3.0 ml of blood were used to compare Aquios CL together with AQUIOS Tetra‐1 Panel against FACSCalibur™ as the reference method. Agreement between the dedicated and the reference method was assessed using the regression analysis, Bland Altman plots and/or Scott percentage. The failure rate was 1.54% and 6.1% in laboratory 1 and 2 respectively. The mean intra‐laboratory variability was 4.1 % for CD4 absolute count and 3.1 % for CD4% at ITM. While at CDC, it was 4.9% and 2.3% for CD4 counts and CD4% respectively. The average inter‐assay variability was 4.2% and 1.6% for CD4 counts and CD4% respectively at ITM while at CDC laboratory it was 5.7% and 2.1% for CD4 counts and CD4% respectively. The mean carry‐over was less than 2% in both laboratories. The observed bias was always within the optimal limits proposed by WHO and the misclassification was less than 3.8% at 350 and 500 CD4/ml levels. Aquios CL is globally easy to use. However, the frequency of notifications of “potential specimen or gating issue” required user intervention for up to 30% of all specimens tested in this evaluation. The manufacturer was informed of this finding.

of 40


Labelling

1. Labels 2. Instructions for use

of 40


1. Labels

of 40

Special Instructions: N/A

Description: CTN; AQUIOS GENERIC BCI BRANDING 2" X 1-3/4" X 3-3/16" LYSING KIT

Ink Color(s): 100% Black, PMS 320C, PMS 368C

Part Number: B28994 Revision: AA Label Size: 2" x 1-3/4" x 3-3/16” Scale: 1 : 1

26°C

18°C

B23538

AQUIOS Lysing Reagent Kit

Intended for preparation of leukocytes from whole blood for flow cytometry/Diseñado en la preparación de leucocitos de sangre total para citometría de flujo./ Предназначено для приготовления лейкоцитов из цельной крови для проточной цитометрии/ Akış sitometrisi için tüm kandan lökosit hazırlama amacını taşır

ライズ試薬キット

体外診断用医薬品

1 x 38 mL Reagent A1 x 15 mL Reagent B

Manufactured forBeckman Coulter Ireland Inc.LismeehanO'Callaghan's MillsCo. Clare, Ireland 353 (0)65 683 1100

ベックマン

有明ウエストタワー東京都江東区有明三丁目製造販売元

© 2013 Beckman Coulter, Inc.Printed in USA Made in USA B28994-AA

: •コールター株式会社 5番7号

TOC

Reagent ASaponin.............< 0.5 g/L Potassium pyro sulfate........< 0.5 g/L

Reagent BSodium sulfate…... < 60 g/LBuffering Agent…...< 4 g/L

Reactive Ingredients:

HOLD AREA for Barcode, Lot and

Exp. symbols1-1/8” x 2-1/2”

of 40

Special Instructions: Barcode size: 3/8” x 3/8”; Hold area size: 1/2” x 1/2”; Location: 5/16” from bottom, 3/8” from left edge.

Description: LBL; PRIM, AQUIOS LYSING REAGENT A Ink Color(s): 100% Black, PMS 368C, PMS 320C

Part Number: B23562 Revision: AA Label Size: 1-1/4" x 2” Scale: 1 : 1

AQUIOS Lysing Reagent A

38 mL

26°C18°C

Printed in USA Made in USA © 2013 Beckman Coulter, Inc.

Manufactured forBeckman Coulter Ireland Inc.Co. Clare, Ireland 353 (0)65 683 1100

B23562-AA

Hold Area for Lot, Exp. and Container no.

Barcode Hold Area

of 40

Special Instructions: Barcode size: 3/8” x 3/8”; Hold area size: 1/2” x 1/2”; Location: 5/16” from bottom, 3/8” from left edge.

Description: LBL; PRIM, AQUIOS LYSING REAGENT B Ink Color(s): 100% Black, PMS 368C, PMS 320C

Part Number: B23563 Revision: AA Label Size: 2" x 1-1/4” Scale: 1 : 1

AQUIOS Lysing Reagent B

15 mL

Printed in USA Made in USA © 2013 Beckman Coulter, Inc. B23563-AA

26°C18°CManufactured for

Beckman Coulter Ireland Inc.Co. Clare, Ireland 353 (0)65 683 1100

Hold Area for Lot, Exp. and Container no.

Barcode Hold Area

of 40

B23565

Ink Color(s): N/ADescription:

Part Number: Revision: Scale: 2:1

LBL; PRI TEM, AQUIOS IMMUNO-TROL LOW CELLS

Reduce 50%, REFERENCE BASE LABEL: B29137

Label Size: 1-3/8" x 1-5/8"

Special Instructions:

AQUIOS IMMUNO-TROL Low Cells

Printed in USAEUH208

© 2014 Beckman Coulter, Inc.

AB

B23565-AB

Made in USA

LOT

of 40

B23566

Ink Color(s): N/ADescription:


LBL; PRI TEM, AQUIOS IMMUNO-TROL CELLS

Reduce 50%, REFERENCE BASE LABEL: B29137

Label Size: 1-3/8" x 1-5/8"


AQUIOS IMMUNO-TROL Cells

Printed in USAEUH208


AB

B23566-AB

Made in USA

LOT

of 40


Ink Color(s): 100% BlackDescription:

B23567

LBL; SEC TEM, AQUIOS IMMUNO-TROL KIT

N/ASpecial Instructions:

AB Label Size: 2-6/8" x 2-6/8"

LOT

of 40


Description: LBL; PRIM, AQUIOS SODIUM HYPOCHLORITE Ink Color(s): 100% Black, PMS 368C, PMS 320C, PMS 186C

Part Number: B23568 Revision: AB Label Size: 2" x 2-3/4" Scale: 1 : 1

Hold Area for Lot and Exp

Printed in USA Made in USA© 2014 Beckman Coulter, Inc.

Manufactured forBeckman Coulter Ireland Inc.Co. Clare, Ireland 353 (0)65 683 1100

50 mL

B23568-AB

26°C

18°C


DANGER H314, 401

of 40

Special Instructions: Barcode size 3/8” x 3/8”, location not critical

Description: AQUIOS Cleaning Agent, Secondary Label Ink Color(s): 100% Black, PMS 320C, PMS 368C, PMS 186C

Part Number: B25694 Revision: AB Label Size: 5-1/4” x 3" Scale: 1 : 1

THIS BOX IS FOR TECH COMM INFORMATIONAL USE ONLY. Refer to “4print.zip” file for all printing specifications.

HOLD AREA forLot, Exp, and Container ID.

1-1/4" x 3"DO NOT VARNISH

2D BarcodeHold AreaDO NOT VARNISH

www.beckmancoulter.com/ifu

Printed in USA Made in USA© 2014 Beckman Coulter, Inc.

Beckman Coulter Ireland Inc.LismeehanO'Callaghan's MillsCo. Clare, Ireland 353 (0)65 683 1100

B25698 500 mL

For use as a cleaning agent for AQUIOS flow cytometer components that come in contact with blood samples./ Para uso como agente limpiador de componentes del citómetro de flujo de AQUIOS que entran en contacto con las muestras de sangre./ Используется в качестве очистителя компонентов AQUIOS проточного цитометра, которые пребывают в контакте с образцами крови./ Kan örnekleriyle temas eden AQUIOS akış sitometresi bileşenleri için temizleme ajanı olarak kullanılır.

May cause allergy or asthma symptoms or breathing difficulties if inhaled. May cause an allergic skin reaction. Avoid breathing vapours. In case of inadequate ventilation, wear respiratory protection. IF INHALED: Remove person to fresh air and keep at rest in a position comfortable for breathing. If experiencing respiratory symptoms: Call a POISON CENTER or doctor/physician. Wear protective gloves, protective clothing and eye/face protection. Take off contaminated clothing and wash it before use.

B25694-AB

26°C

18°C

REACTIVE INGREDIENTS: A solution of a proteolytic enzyme/反応成分: 蛋白質分解酵素溶液

AQUIOS Cleaning Agent

製造販売元: ベックマン·コールター株式会社東京都江東区有明三丁目5番7号TOC有明ウエストタワー

DANGER Subtilisin <1%reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC# 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC# 220-239-6](3:1) <0.05%

of 40

Special Instructions: NoneDescription: Carton, Preprint, AQUIOS IMMUNO-TROL Cells and AQUIOS IMMUNO-TROL Low Cells and AQUIOS IMMUNO-TROL Cells (PLG/Tetra) and AQUIOS IMMUNO-TROL Low Cells (PLG/Tetra)

Ink Color(s): 100% Black, PMS 320C, PMS 368C

Part Number: B28990 Revision: AB Label Size: 3-15/16" x 1-9/32” x 4-9/16” Scale: 1 : 1


2-3/4” x 2-3/4”Sec Label Area

Intended as an assayed, lysable whole blood quality control product for immunophenotyping analysis using monoclonal antibody reagents for flow cytometry./ Para uso como control de calidad analizado a base de sangre total lisable para análisis de inmunofenotipificación utilizando reactivos de anticuerpos monoclonales para citometría de flujo./ Анализированноый и лизированный контроль из цельной крови, предназначенный для определения иммунофенотипа с помощью реагентов, содержащих моноклональные антитела, для проточной цитометрии./ Akış sitometresi için monoklonal antikor reaktifler kullanan immunfenotiplendirme analizi için test edilmiş ve lizabl tam kan kalitesi kontrol ürünü olarak tasarlanmıştır.

Potential Biohazardous Material

Printed in USA Made in USA© 2014 Beckman Coulter, Inc.B28990-AB

製造販売元: ベックマン·コールター株式会社東京都江東区有明三丁目5番7号TOC有明ウエストタワー

Beckman Coulter Ireland Inc.LismeehanO’Callaghan’s MillsCo. Clare, Ireland 353 (0)65 683 1100

www.beckmancoulter.com/ifu

of 40

Hold area for Barcode, Lot & Exp.

4” x 4”

Hold

area

for B

arco

de,

Lot &

Exp

. 4”

x 4”

Special Instructions: Overlay of colors resulting in new colors is acceptable.

Description: AQUIOS SHEATH SOLUTION Ink Color(s): GCMI #90, GCMI #387, GCMI #2081

Part Number: B28991 Revision: AA Label Size: N/A Scale: 1 : 1


シース液

AQU

IOS

Sheath Solutionシース液

AQU

IOS Sheath Solution

AQU

IOS

Shea

th S

olut

ion

シース液

REACTIVE INGREDIENTS:/ Sodium Chloride/ ......…

….…

…..........7.93 g/L

Disodium EDTA/ ...……

…..............0.38 g/L

Potassium Chloride/ ..........……

……

.......0.40 g/LMonosodium Phosphate/ …

..0.19 g/LDisodium Phosphate/ .…

........1.95 g/LSodium Fluoride/ ..…

……

…............0.30 g/L

塩化ナトリウム

二ナトリウムEDTA

塩化カリウム

一ナトリウムリン酸塩

二ナトリウムリン酸塩

フッ化ナトリウム

INTENDED USE: A non-fluorescent, balanced electrolyte solution for use on AQUIOS Flow Cytometers with light scatter and fluorescent applications./ APLICACIÓN: Una solución electrolítica balanceada, no fluorescente para uso en los Citómetros de Flujo AQUIOS con aplicaciones de dispersión de luz y fluorescencia./ ПРЕДНАЗНАЧЕНИЕ: Нефлуоресцирующий сбалансированный раствор электролита, предназначенный для использования в проточных цитометрах типа AQUIOS с рассеиванием света и флюоресценцией./ KULLANIM AMACI: Işık saçılımı ve floresan uygulamalı AQUIOS Akış Sitometrelerinde kullanılma amacını taşıyan, floresan olmayan dengeli bir elektrolit çözeltisidir.

B28991-AA

B25697

B256

97

Beck

man C

oulte

r Ire

land I

nc.

Lisme

ehan

O’Ca

llagh

an’s

Mills

Co. C

lare,

Irelan

d 353

(0)6

5 683

1100

Print

ed in

USA

Mad

e in U

SA©

2013

Bec

kman

Cou

lter,

Inc.

製造販売元: ベックマン·コールター株式会社

東京都江東区有明三丁目5番7号

TOC有明ウエストタワー

10 L

成分:


26°C

18°C

ベックマン・コールター株式会社

of 40


Description: CTN; AQUIOS GENERIC BRANDING 2” X 1-1/4” X 2-3/4” Ink Color(s): 100% Black, PMS 320C, PMS 368C

Part Number: B28992 Revision: AB Label Size: 2" x 1-1/4" x 2-3/4” Scale: 1 : 1

B28992-AB


Beckman Coulter Ireland Inc.LismeehanO'Callaghan's MillsCo. Clare, Ireland 353 (0)65 683 1100

Reactive Ingredient: Murine Monoclonal Antibody 0.9 mL成分：マウスモノクローナル抗体 0.9 mL

製造販売元: ベックマン•コールター株式会社東京都江東区有明三丁目5番7号TOC有明ウエストタワー


8°C

2°C

For Use In Flow Cytometric Analysis/Para uso en análisis por citometría de flujo/Для использования в проточном цитометрическом анализе/Akış Sitometrisi Analizinde Kullanmak İçindir

HOLD AREA FOR LABEL3” x 2-1/2”

of 40


Description: CTN; AQUIOS GENERIC BRANDING 4-3/16" x 2-3/16" x 4-3/8” SODIUM HYPOCHLORITE

Ink Color(s): 100% Black, PMS 320C, PMS 368C, PMS 186C

Part Number: B28993 Revision: AB Label Size: 4-3/16" x 2-3/16" x 4-3/8” Scale: 1 : 1

HOLD AREA for Barcode, Lot and

Exp.


B28993-AB

Manufactured forBeckman Coulter Ireland Inc.LismeehanO'Callaghan's MillsCo. Clare, Ireland 353 (0)65 683 1100

Printed in USA Made in USA

製造販売元: ベックマン•コールター株式会社東京都江東区有明三丁目5番7号TOC有明ウエストタワー

AQUIOS Sodium Hypochlorite Solution is a cleaning agent used with AQUIOS flow cytometers. The solution maintains instruments in optimal condition./La solución de hipoclorito de sodio AQUIOS es un agente de limpieza utilizado con los citómetros de flujo AQUIOS. La solución mantiene el instrumento en óptimo estado./Раствор гипохлорита натрия AQUIOS является очищающим средством, которое используется для проточных цитометров AQUIOS. Раствор поддерживает работоспособность приборов в оптимальных условиях./AQUIOS Sodyum Hipoklorit Solüsyonu AQUIOS akış sitometreleriyle kullanılan bir temizlik ajanıdır. Solüsyon aletleri optimum durumda tutar.


次亜塩素酸ナトリウム溶液AQUIOS Sodium Hypochlorite Solution

B23536

4 x 50 mL

Reactive Ingredients: Sodium Hypochlorite 5.25% Surfactant成分: 次亜塩素酸ナトリウム 5.25% 界面活性剤

DANGER H314, 401Sodium Hypochlorite 1-10%

26°C

18°C

of 40



B28995

LBL; SEC TEM, AQUIOS TETRA-1 KIT


Label Size: 2-3/8" x 2-7/8"

LOT

Made in USA

EUH208


50 Tests

B23533

CD4-RD1

REF

Iodoacetamide <0.1%

B28995-AB

IVD

AB

CD45-FITC

CD3-PC5CD8-ECD

Printed in USA

AQUIOS Tetra-1 Panel

of 40



B29060

LBL; SEC TEM, AQUIOS TETRA-2+ KIT


Label Size: 2-3/8" x 2-7/8"

LOT

Made in USA

EUH208


50 Tests

B23534

(CD56+CD16)-RD1

REF

Iodoacetamide <0.1%

B29060-AB

IVD

AB

CD45-FITC

CD3-PC5CD19-ECD

Printed in USA

AQUIOS Tetra-2+ Panel

of 40

IVD

B25700

AQUIOS IMMUNO-TROL Low Cells

REF

B29067-AB

of 40

Special Instructions: Barcode size 2-3/8 x 1-1/8; Hold area location: Left edge; Label folds at barcode hold area.

Ink Color(s): 100% Black, PMS 368C, PMS 320C, PMS 150C

Part Number: B29133 Revision: AA Label Size: 2-3/8” x 2-7/8” Scale: 1 : 1

Description: LBL; SEC, AQUIOS Tetra KIT Base Label

Hold area for Barcode,

Lot and Exp

50 Tests

HarmfulR22,S28

Content: 1 x 0.9 mL, cap/ bouchon/ cappuccio/ Deckel/ tapón/ tampa/ hætte/ lock/ krytka/ zatyczka/ крышка/ kapak/ hette/ キャップ/ krytka/ kapica/ kaas.

B291

33-A

A

of 40


Description: AQUIOS IMMUNO-TROL Primary Base Label Ink Color(s): 100% Black, PMS 368C, PMS 320C

Part Number: B29137 Revision: AA Label Size: 1-21/32" x 1-3/8” Scale: 1 : 1

Hold area for barcode lot and exp.

8°C

2°C

3 mL

Beckman Coulter Ireland Inc.Co. Clare, Ireland 353 (0)65 683 1100

< 0.1% NaN3

B291

37-A

A

of 40



B29140

LBL; SEC TEM, AQUIOS IMMUNO-TROL LOW KIT


AB Label Size: 2-6/8" x 2-6/8"

LOT

of 40

IVD B29143-AB

B23535REF

AQUIOS IMMUNO-TROL Cells

of 40

Description: LBL; PRIM, TEM, AQUIOS Tetra-1 Panel Ink Color(s): 100% Black, PMS 320C

Part Number: B38717 Revision: AB Label Size: 1-3/4” x 1” (overlap 1/8”) Scale: 1 : 1

Special Instructions: Lot and Exp hold area: not critical. Container: unique ID number, 00001 - NNNNN, where the maximum for NNNNN is 16383.

8°C2°C

0.9 mL50 tests

B387

17-A

B

AQUIOS Tetra-1 PanelCD45-FITCCD4-RD1CD8-ECDCD3-PC5

Beckman Coulter Ireland Inc.Co. Clare, Ireland 353 (0)65 683 1100 Printed in USA Made in USA © 2014 Beckman Coulter, Inc.

EUH208

Hold area for Lot, Exp, and C:

of 40

Description: LBL; PRIM, TEM, AQUIOS Tetra-2+ Panel Ink Color(s): 100% Black, PMS 320C

Part Number: B38718 Revision: AB Label Size: 1-3/4” x 1” (overlap 1/8”) Scale: 1 : 1

Special Instructions: Lot and Exp hold area: not critical. Container: unique ID number, 00001 - NNNNN, where the maximum for NNNNN is 16383.

Hold area for Lot, Exp, and C:

8°C2°C

0.9 mL50 tests

B387

18-A

B

CD45-FITC(CD56+CD16)-RD1CD19-ECDCD3-PC5

AQUIOS Tetra-2+ Panel

Beckman Coulter Ireland Inc.Co. Clare, Ireland 353 (0)65 683 1100 Printed in USA Made in USA © 2014 Beckman Coulter, Inc.

EUH208

of 40


2. Instructions for use

of 40

1 of 5

AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/ CD8-ECD/CD3-PC5

B23533 - 50 tests

AQUIOS Tetra-2+ Panel CD45-FITC/ (CD56+CD16)-RD1/ CD19-ECD/CD3-PC5

B23534 - 50 tests

PN B25337-AC

CD45-FITC CD4-RD1 CD8-ECD CD3-PC5 (CD56+CD16)-RD1 CD19-ECD

Specificity CD45 CD4 CD8 CD3 CD56, CD16 CD19

Clone B3821F4A SFCI12T4D11 SFCI21Thy2D3 UCHT1 N901/NKH-1, 3G8 J3-119

Hybridoma NS-1 x BALB/c NS-1 x BALB/c NS-1 x BALB/c NS-1 x BALB/c NS-1 x BALB/c, SP2/0 x BALB/c

NS-1 x BALB/c

Immunogen Transfectant containing cDNA of human CD45

Human peripheral T lymphocytes

Human thymocytes Human thymocytes and peripheral blood lymphocytes from a person with Sezary cell leukemia

Human chronic myeloid leukemia cells, Human neutrophils

SKLY 18 lymphoma cells

Ig Chain IgG2b IgG1 IgG1 IgG1 IgG1 (both) IgG1

Species Mouse Mouse Mouse Mouse Mouse (both) Mouse

Source Ascites fluid Conditioned media Conditioned media Conditioned media Ascites fluid, Conditioned Media

Conditioned media

Purification Affinity chromatography

Affinity chromatography



Affinity chromatography (both)


Fluorescence Excites at 468-509 nm Emits at 504-541 nm

Excites at 486-580 nm Emits at 568-590 nm



Excites at 486-580 nm Emits at 568-590 nm (both)


Conjugation FITC (Fluorescein Isothiocyanate)

RD1 (Phycoerythrin) ECD (Phycoerythrin -Texas Red-X)

PC5 (Phycoerythrin-Cy5)

RD1 (Phycoerythrin) (both)

ECD (Phycoerythrin -Texas Red-X)

Molar Ratio FITC/Protein: 3-10 RD1/Protein: 0.5-1.5 ECD/Protein: 0.5-1.5 PC5/Protein: 0.5-1.5 RD1/Protein: 0.5-1.5 (both)

ECD/Protein: 0.5-1.5

MONOCLONAL ANTIBODY For In Vitro Diagnostic Use

Rx Only

INTENDED USE AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel monoclonal antibody reagents are for use on the AQUIOS CL Flow Cytometer with peripheral whole blood for immunophenotyping. These reagents are indicated for use in the immunologic assessment of patients having, or suspected of having, immune deficiency. These reagents provide identification and enumeration of:

� AQUIOS Tetra-1 Panel Monoclonal Antibody Reagents � Total CD3+, CD3+CD4+, CD3+CD8+,

CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts.

� CD45+ absolute count

� CD45+ Low SS (lymphocytes) percentage and absolute count

� AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagents � Total CD3+, CD3-CD19+, CD3-CD56+ and/or

CD16+ lymphocyte percentages and absolute counts.

� CD45+ absolute count

� CD45+ Low SS (lymphocytes) percentage and absolute count

Refer to the AQUIOS Tetra System Guide (PN B26364) for instructions on how to use these reagents in the system and their respective Performance Characteristics.

SUMMARY AND EXPLANATION The leukocyte common antigen (CD45) is a transmembrane-type protein and expressed at high levels on nucleated hematopoietic cells with the exclusion of megakaryocyte/platelet and erythroid series.61 CD45-assisted Leukocyte gating along with CD4 allow readily enumeration of absolute count and percentages of CD4 T cells.62,63,64 The expression of CD45 density is useful for discriminating between normal and malignant leucocytes cells. The density of

expression of CD45 is weak in some malignant cells (i.e. acute myeloid leukemias) thus, enabling malignant cells to be distinguished from normal ones. The lymphocyte population of human peripheral blood is composed of three cell types: T (thymus-derived), B (bone marrow-derived), and NK (Natural Killer) cells.4 These cell types are morphologically indistinguishable by microscopy but can be identified by characteristic antigenic differences in their cell membranes. T, B, and NK lymphocytes play central roles in immune system function. Different subtypes of T lymphocytes may recognize specific antigens, execute effector functions and/or control both the type and intensity of cellular and/or humoral immune responses. Upon activation by antigens or macrophages via T lymphocytes, specific B lymphocytes differentiate into plasma cells which produce and secrete specific immunoglobulins (Ig). NK lymphocytes have been identified as a discrete population of cytolytic effectors and appear to play an integral part in regulation of hematopoiesis, the defense against viral infection, and the destruction of malignant tumor cells. The NK lymphocyte mediated cytolytic activity occurs without restriction by class I or II major histocompatibility complex (MHC) antigens.4

CLINICAL RELEVANCE CD3+, CD4+, CD8+, and/or CD19+ Lymphocytes CD3+, CD4+, CD8+, and/or CD19+ lymphocyte percentages and absolute counts may be used as aids to evaluate immune competency underlying known or unknown disease states and to monitor lymphocyte levels following organ transplantation.17,25,26,27,28,29,30,31,32,33

To illustrate, identification of abnormal levels of CD3+, CD4+, CD8+, and/or CD19+ lymphocytes may aid in the diagnosis and/or prognosis of unidentified disease conditions in patients with low white blood cell counts. Altered percentages of CD3+, CD4+, CD8+, and/or CD19+ lymphocytes recorded following organ (kidney, heart, liver, lung) transplantation suggest T (CD3+, CD4+, CD8+) and/or B (CD19+) lymphocyte measurements may be useful as aids in monitoring these cellular populations.

Identification of abnormal levels of CD4+ immunodeficiency, and corresponding CD4+/CD8+ ratios, might also aid in the diagnosis and/or prognosis of immunodeficiency disease. For example infection with human immunodeficiency virus (HIV), the etiologic agent of acquired immunodeficiency syndrome (AIDS), results in profound immunosuppression due predominantly to a selective depletion of the CD4+ lymphocytes that express the receptor for the virus.17,34

Progressive clinical and immunologic deterioration generally correlates with a decreasing CD4+ lymphocyte count.17

CD3-(CD56+CD16)+ Lymphocytes NK lymphocyte populations have been functionally defined as a lymphocyte population capable of mediating non-MHC restricted cytotoxicity against targets such as certain tumor and virus-infected cells.35

CD4/CD8 Ratio Disease-related changes in CD4+ and/or CD8+ lymphocyte levels might alter CD4/CD8 inducer suppressor/cytotoxic cell ratios. As a result, CD4/CD8 ratios might be useful as diagnostic and/or prognostic indicators of immune competence. CD4/CD8 ratios in conjunction with CD4+ lymphocyte cell numbers have been the most widely used laboratory parameters for evaluation of AIDS-related complex and AIDS.17,36 CD4/CD8 ratios approach zero in advanced AIDS patients with no detectable levels of CD4+ lymphocytes.17 In such cases, CD8+ lymphocyte levels might be normal, increased or decreased. Decreased CD4+ and CD8+ lymphocyte percentages without significant changes in CD4/CD8 ratios have been observed in patients with stable renal allograft function after transplantation.5 In addition, low CD4/CD8 ratios and decreased percentages of CD4+ lymphocytes have been documented in patients during phenotypic reconstitution following purged autologous bone marrow transplantation.31,32

CD45+ Cells The CD45+ cells are identified as white blood cells (leukocytes) since CD45 antigen is expressed on every type of hematopoietic cell except mature erythrocytes and their immediate progenitors7,8,9,10 and is not identified on cells of non-hematopoietic origin. CD45 in combination with Side Scatter may be used to

of 40

2 of 5

define leukocytes and to differentiate discrete cell populations for immunophenotyping.

CD45+ Low SS (Lymphocytes) The CD45+ Low SS cells are identified as the lymphocyte population. The lymphocyte population can be further differentiated into discrete cell populations for immunophenotyping.

Lymphocyte Immunophenotyping Panel AQUIOS Tetra-2+ Panel provides the ability to enumerate an individual's major lymphocyte subsets: T, B and NK. The reagent can function as a quality control check (LymphoSum) to estimate CD45/SS Lymphocyte gate recovery and account for all Lymphocyte subsets. Total lymphocyte percentage should be determined using the following formula:

Total Lymphocyte Percentage (%) = %CD3+(T) + %CD19+(B) + %CD3-(CD56+CD16)+ (NK).46,47

Used as a panel, AQUIOS Tetra-1 Panel also functions as a quality control check for a specimen in terms of total lymphocyte percentage and CD3+ lymphocyte measurements across the panel (CD3+ Intrapanel Check).47

PRINCIPLES OF TEST This test depends on the ability of a monoclonal antibody to bind to the surface of cells expressing discrete antigenic determinants. Specific cell staining is accomplished by incubating whole blood with the monoclonal antibody reagent. The AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel monoclonal antibody reagents are each a combination of four or five murine monoclonal antibodies, each conjugated to a specific fluorochrome and specific for different cell surface antigens. Red blood cells are lysed with the AQUIOS Lysing Reagent Kit. The remaining white blood cells are analyzed by flow cytometry using lymphocyte gates. In the first histogram for either reagent, the lymphocyte gate is identified as having bright CD45+ FITC fluorescence and low Side Scatter (SS). For AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/ CD8-ECD/CD3-PC5 the following histograms/plots are used to determine the percentage of positively stained cells: CD3+ (positive PC5 fluorescence only), CD3+CD4+ (positive PC5/RD1 fluorescence), CD3+CD8+ (positive PC5/ECD fluorescence). For AQUIOS Tetra-2+ Panel CD45-FITC/ (CD56+CD16)-RD1/ CD19-ECD/CD3-PC5, the following histograms/plots are used to determine the percentage of positively stained cells: CD3+ (positive PC5 fluorescence only), CD3-CD56+ and/or CD16+ (positive RD1 fluorescence only), CD3-CD19+ (positive ECD fluorescence only). The AQUIOS System Software with Tetra-1 and Tetra-2+ tests in conjunction with

AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel monoclonal antibody reagents provides automated analysis of lymphocyte subpopulations. (Refer to the AQUIOS Tetra System Guide PN B26364 for more details.)

REAGENTS See table on page 1.

REAGENT CONTENTS The antibody concentration is 1.445/0.182/0.365/ 0.365 µg/test for CD45-FITC/CD4-RD1/CD8-ECD/ CD3-PC5 and 1.445/0.022/0.091/0.247/0.365 µg/test for CD45-FITC/ CD56-RD1/CD16-RD1/CD19-ECD/ CD3-PC5.

The concentration of nonantibody reagents is 0.2% BSA, 0.01 M potassium phosphate, 0.15 M NaCl, 0.1% NaN3 and stabilizers.

STATEMENT OF WARNINGS

Iodoacetamide <0.1%

May produce an allergic reaction.

Safety Data Sheet is available at techdocs.beckmancoulter.com.

1. These reagents contain 0.1% sodium azide. Sodium azide under acid conditions yields hydrazoic acid, an extremely toxic compound. Azide compounds should be flushed with running water while being discarded. These precautions are recommended to avoid deposits in metal piping in which explosive conditions can develop. If skin or eye contact occurs, wash excessively with water.

2. Specimens, samples, and all material coming in contact with them should be handled as if capable of transmitting infection and disposed of with proper precautions.

3. Do not remove the supplied Cap with Septa from the packaging until ready to use.

4. Ensure that the shipping cap has been replaced with the Cap with Septa immediately prior to loading on the system. Otherwise, damage will occur to flow cytometer system.

5. Never pipette by mouth and avoid contact of samples with skin and mucous membranes.

6. Do not use reagent beyond the expiration date on the vial label.

7. Minimize exposure of reagent to light during storage or incubation.

8. Avoid microbial contamination of reagent or erroneous results may occur.

9. Use Good Laboratory Practices (GLP) when handling reagent.

10. Review all data plots and histograms before reporting results. Refer to the AQUIOS Tetra System Guide (PN B26364) for the Flow Cytometry Gating Strategy and Data Plot Examples.

11. Harmful if swallowed. 12. After contact with skin wash immediately with

plenty of water.

13. Do not operate the system with an uncapped vial of antibody reagent.

14. Once the cap with septa has been pierced the vial has the potential to leak if the bottle is not upright.

STORAGE CONDITIONS AND STABILITY Unopened reagent is stable to the expiration date on the vial label when stored at 2-8°C. Opened vials are stable for 90 days when stored at 2-8°C. Return reagent to 2-8°C immediately after use. Do not freeze. Minimize exposure to light. Monoclonal antibody reagent vials may be left at room temperature on board the AQUIOS System for up to a maximum of 80 total cumulative hours. If the facility operates the System for periods of longer than 8 continuous hours, it is suggested that the carousel with the monoclonal antibody vials be stored at 2-8°C and a second carousel with monoclonal antibody vials be loaded for the next period of time.

EVIDENCE OF DETERIORATION Any change in the physical appearance of these reagents (normal appearance is a clear, pink liquid) or any major variation in values obtained for control samples may indicate deterioration and the reagent should not be used.

REAGENT PREPARATION No preparation is necessary. These monoclonal antibody reagents are used directly from the vial. Bring reagent to 18-26°C prior to use.

NOTE: Ensure the shipping cap has been removed and replaced with the Cap with Septa provided in the packaging before placing vial in system carousel.

SPECIMEN COLLECTION � Refer to the AQUIOS Tetra System Guide

(PN B26364) for the minimum volume of blood required for analysis.

� Optimal staining is achieved with white blood cell counts in the range of 0.3-25 x 103 cells/µL.

� For detailed information on the collection of whole blood by venipuncture and interfering conditions, refer to Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture.38

1. Collect venous blood specimen aseptically by

venipuncture into a blood collection tube using the recommended anticoagulant EDTA.

2. Perform a total white blood cell count using an established procedure.

PROCEDURE FOR IMMUNOFLUORESCENCE CELL SURFACE STAINING WITH TETRA-1 PANEL AND TETRA-2+ PANEL

MATERIAL SUPPLIED AQUIOS Tetra-1 Panel

B23533 - 50 tests (0.9 mL) AQUIOS Tetra-2+ Panel

B23534 - 50 tests (0.9 mL)

B30104 – Cap with Septa (1)

MATERIALS REQUIRED BUT NOT SUPPLIED AQUIOS Lysing Reagent Kit, PN B23538 –100 tests

AQUIOS IMMUNO-TROL Cells, PN B23535

OR

AQUIOS IMMUNO-TROL Low Cells, PN B25700

Blood collection tubes with EDTA anticoagulant

AQUIOS CL Flow Cytometer

PROCEDURE 1. Bring the antibodies to 18-26°C.

2. If the vial has not been previously used on the system, remove the shipping cap and replace with the Cap with Septa provided in the packaging. Replacement of the shipping cap should only occur immediately prior to loading on the system to allow the system to monitor and accurately reflect the open vial stability claim.

NOTE: Once Cap with Septa is placed on vial do not remove.

3. LOAD the sample on the AQUIOS system. Select “Patient” in the “Test Request”.

� For autoloader, insert tube(s) into the instrument cassette and place the cassette on the system.

� For single tube loader, mix the sample immediately before placing the tube in the tube sampling area. Scan the tube and place on the system.

4. GO is automatically initiated when sample(s) are placed on the system.

� For autoloader, the system runs the sample(s) when the cassette is loaded.

of 40

3 of 5

� For single-tube loader, the system runs the sample(s) when the door is closed.

5. Allow the system to prepare the sample(s) and to analyze.

Refer to the AQUIOS Tetra System Guide PN B26364 for detailed instructions.

QUALITY CONTROL PROCEDURE Daily Quality Control is a critical component of ensuring the system’s performance for the Tetra application. Refer to the AQUIOS Tetra System Guide (PN B26364).

LIMITATIONS 1. The reagents are for use only on AQUIOS Flow

Cytometers.

2. Specimens must be prepared with AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel reagents and AQUIOS Lysing Reagent System within 24 hours of collection.

3. The CD45+ absolute count and CD45+ Low SS percentage and absolute counts should only be used for Immunophenotyping flow cytometric analysis.

4. Retain specimens in blood collection tubes at room temperature prior to staining and analyzing.

5. Do not refrigerate specimens. Refrigerated specimens may give aberrant results.

6. The recommended cell viability for venous blood specimens is >90%.

7. AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel monoclonal antibody reagents are designed for use with whole blood samples. They may also be used with AQUIOS IMMUNO-TROL Cells, and/or AQUIOS IMMUNO-TROL Low Cells. The reagents are not recommended for use with fresh or frozen mononuclear cell preparations.

8. Do not dilute, aliquot, or freeze the reagents. The product should be used according to labeled instructions.

9. In patients treated with anti-human monoclonal antibody therapies, detection of the specific targeted antigens may be diminished or absent due to partial or complete blocking by the treatment antibody.41,42,43

10. Abnormal states of health are not always represented by abnormal percentages of certain leukocyte populations. An individual in an abnormal state of health may show the same leukocyte percentages as a healthy person. Use test results in conjunction with clinical and other diagnostic data.

11. Certain patients may present special problems due to altered or very low numbers of certain cellular populations.

12. Patients with chronic HIV or elevated viremia may exhibit lower than expected NK lymphocyte results due to a phenotypic shift in the NK cell subsets. In these clinical conditions there is a selective loss of CD56dimCD16+ NK cell subset and an expansion of other pathologic NK cell populations. Use test results in conjunction with clinical and other diagnostic data.44,45

13. Results obtained with flow cytometry may be erroneous if the laser is misaligned or the gates and regions are improperly set.

14. In some specimens, purity of the lymphocyte region may be decreased due to non-lymphoid contaminants with low SS and high CD45 fluorescence similar to lymphocyte populations. These samples may meet CD3+ Reliability Check acceptance criteria, as the relative proportion of CD3+CD4+ and CD3+CD8+ cells remains constant, yet the result may not be accurate. A review of all data plots for the presence of the

expected staining patterns is recommended for all samples.

15. Performance has not been established for pediatric use.

PERFORMANCE CHARACTERISTICS Refer to the AQUIOS Tetra System Guide (PN B26364) for information on Reference Ranges, Linearity, Accuracy of Method, Precision, Analytical Measuring Ranges and Quality Control.

SPECIFICITY The CD45 antigen is expressed on every type of hematopoietic cell except mature erythrocytes and their immediate progenitors.7,8 It has not been detected in differentiated nonhematopoietic tissue.7,8,9,10 The CD3 antigen is normally present on the cell surface of mature thymocytes and resting and activated peripheral blood mature T lymphocytes (both inducer and suppressor/cytotoxic populations).12,13,14

The CD4 antigen is present on thymocytes and the inducer T lymphocyte population in peripheral blood.13,14 It is also expressed at low density on monocytes.17 The CD8 antigen is normally present on approximately 80% of thymocytes and approximately 30-35% of peripheral blood T lymphocytes and some natural killer cells.15,18 The CD16 antigen is the low-affinity receptor for IgG (FcγRIII) that binds immune complexes, but not monomeric IgG. The CD16 antigen exists in two different forms encoded by two different genes: FcγRIIIA (or III-2) and FcγRIIIB (or III-1). The genetic heterogeneity of CD16 generates alternative membrane-anchored molecules. One is a transmembrane form (FcγRIIIA, 50 – 65 kDa) expressed on NK cells, monocytes and macrophages. The other is a glycosylphosphatidylinositol (GPI)-anchored form (FcγRIIIB, 48 kDa) only expressed on neutrophils.52,53 It has been shown that the CD16 antigen can be non-covalently associated within the membrane of NK cells, to the 16 kDa CD3ζ chain,54 or to the dimeric FcRγ chain.55 The 3G8 monoclonal antibody (mAb) binds to FcγRIIIA as well as to FcγRIIIB (strongly). It was shown to block almost completely the binding of IgG dimers to FcγRIIIB.56 Experiments where amino acid mutations were made to the FcγRIIIB molecule showed that the 3G8 mAb is affected by Lys162 and Val164 substitutions in the FG loop of the membrane-proximal Ig-like domain of the molecule.57,59 The 3G8 mAb has been assigned to the CD16 cluster of differentiation at the Fifth International Workshop on Human Leucocyte Differentiation Antigens held in Boston, USA, in 1993.58

The CD19 antigen is expressed on all B cells, including early progenitor B cells.3 It can also be found on follicular dendritic cells and myelomonocytic lineage progenitor cells, but is not expressed on T cells, monocytes or granulocytes.1,19,20 The CD56 antigen is expressed on a subpopulation of lymphocytes that demonstrate natural killer (NK) activity (and also on various types of non-circulating cells of neural and/or neuroendocrine origin). This subpopulation consists of both natural killer cells (CD3-CD56+) and a subset of T cells (CD3+CD56+).1,19,21,23

CD3-CD56+ cells are capable of mediating non-TCR mediated cytotoxicity in peripheral blood.21,22 CD56 is not expressed on other T or B lymphocyte, monocyte, granulocyte or erythrocyte populations.23,24,25

The antigen specificity of the CD45, CD3, CD4, and CD8 monoclonal antibodies comprising the AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/ CD8-ECD/CD3-PC5 monoclonal antibody reagent has been previously established by the First and the Fifth International Workshops for Leukocyte Typing.1,2 The antigen specificity of the CD45, CD3, CD19, CD56 and CD16 monoclonal antibodies comprising the AQUIOS Tetra-2+ Panel CD45-FITC/CD56-RD1/ CD19-ECD/CD3-PC5 CD16 monoclonal antibody reagent has been previously established by the First, Fourth and Fifth International Workshops for Leukocyte Typing.6,11,16,39

To assess cellular cross-reactivity, the CD3, CD4, CD8, CD19, and CD56 monoclonal antibodies comprising the AQUIOS Tetra-1 Panel CD45-FITC/ CD4-RD1/CD8-ECD/CD3-PC5 and AQUIOS Tetra-2+ Panel CD45-FITC/(CD56+CD16)-RD1/ CD19-ECD/ CD3-PC5 monoclonal antibody reagents were screened on normal human adult donor blood samples. Results consistently demonstrated that the CD3, CD4, CD8, CD19, and CD3-(CD56+CD16)-RD1 antibodies reacted specifically with the appropriate lymphocyte populations. Monocytes were dimly stained with CD4 monoclonal antibody.

REFERENCES 1. Schlossman SF, Boumsell L, Gilks W, Harlan JM,

Kishimoto T, Morimoto C, Ritz J, Shaw S, Silverstein R, Springer R, Tedder TF and Todd RF, eds. Leukocyte Typing V. Oxford, UK: Oxford University Press, 1995, Vol. 1. pp. 262, 263, 268, 270, 507-511; Vol. 2. pp. 1398-1400.

2. Bernard A, Boumsell L, Dausset J, Milstein C and Schlossman SF, eds. Leukocyte Typing. Springer-Verlag, New York; 1984, pp. 28, 41-42, 44,196.

3. McMichael AJ, ed. Leukocyte Typing III. Oxford: Oxford University Press, 1987, pp. 38, 40, 42, 43, 116, 167, 170-172, 176, 199, 202, 206, 302-308, 315, 475, 796-801.

4. Robertson MJ, Cochran KJ, Cameron C, Le J-M, Tantravahi R, and Ritz J. Characterization of a cell line, NKL, derived from an aggressive human natural killer cell leukemia. Experimental Hematology, 1996, 24:406-415.

5. Foon KA and Todd RF. Immunologic classification of leukemia and lymphoma. Blood, 1986, 68:1-31.

6. Newman, W Targan SF, and Fast LD. Immunobiological and immunochemical aspects of the T-200 family of glycoproteins. Mol Imm, 1984, 21:1113-1121.

7. Coffman RL and Weissman IL. B220: A B cell specific member of the T-200 glycoprotein family. Nature, 1981, 289:681-693.

8. Dalchau R and Fabre JW. Identification with a monoclonal antibody of predominantly B lymphocyte specific determinant of the human leucocyte common antigen. J Exp Med, 1980, 153:753-757.

9. Omary MB, Trowbridge IS and Battifora HA. Human homologue of murine T-200 glycoprotein. J Exp Med, 1980, 152:842-852.

10. Dalchau R, Kirkley J and Fabre JW. Monoclonal antibody to a human leukocyte-specific membrane glycoprotein probably homologous to the leukocyte common (L-C) antigen of rat. Eur J Immunol, 1981, 10:737-744.

11. Knapp W, Dörken B, Gilks WR, Rieber EP, Schmidt RE, Stein H, and von dem Borne AEGKr., eds. Leukocyte Typing IV. White Cell Differentiation Antigens. Oxford, UK: Oxford University Press, 1989, pp. 536, 541, 699-702.

12. Reinherz EL and Schlossman SF. The differentiation and function of human T lymphocytes. Cell, 1980, 19:821-827.

of 40

4 of 5

13. Reinherz EL, Meuer S, Fitzgerald KA, Hussey RE, Levine H and Schlossman SF. Antigen recognition by human T lymphocytes is linked to surface expression of the T3 molecular complex. Cell, 1982, 30:735-743.

14. Meuer SC, Acuto O, Hussey RE, Hodgdon JC, Fitzgerald KA, Schlossman SF and Reinherz EL. Evidence for the T3-associated 90K heterodimer as the T-cell antigen receptor. Nature, 1983, 303:808-810.

15. Reinherz EL, Meuer SC, and Schlossman SF. The delineation of antigen receptors on human T lymphocytes. Immunology Today, 1983, 4:5-8.

16. Ravetch, J.V., Perussia, B., "Alternative membrane forms of FcγRIII (CD16) on human natural killer cells and neutrophils. Cell type-specific expression of two genes that differ in single nucleotide substitutions." 1989, J. Exp. Med., 170(2), 481-497.

17. de Martini RN and Parker JW. Immunologic alterations in human immunodeficiency virus infection: A review. J Clin Lab Anal, 1989, 3:56-70.

18. Caligiuri M, Murray C, Buchwald D, Levine H, Cheney P, Peterson D, Komaroff AL and Ritz J. Phenotypic and functional deficiency of natural killer cells in patients with chronic fatigue syndrome. J Immunol, 1987, 139:3306-3313.

19. Barclay AN, Birkeland ML, Brown MH, Beyers AD, Davis SJ, Samoza C, Williams AF eds. The Leucocyte Antigen Facts Book. London: Academic Press, 1993, pp. 106-109.

20. Tedder TF and Isaacs CM. Isolation of a cDNA encoding the CD19 antigen of human and mouse B lymphocytes: a new member of the immunoglobulin superfamily. J Immunol, 1989, 143:712-717.

21. Griffin JD, Hercend T, Beveridge RP, and Schlossman SF. Characterization of an antigen expressed by human natural killer cells. J Immunol, 1983, 130:2947-2951.

22. Hercend T, Griffin JD, Bensussan A, Schmidt RE, Edson MA, Brennan A, Marray C, Daley JF, Schlossman SF, and Ritz J. Generation of monoclonal antibodies to a human natural killer clone. Characterization of two natural killer-associated antigens, NKH1A and NKH2, expressed on subsets of large granular lymphocytes. J Clin Invest, 1985, 75:932-943.

23. Schmidt RE, Murray C, Daley JF, Schlossman SF, and Ritz J. A subset of natural killer cells in peripheral blood displays a mature T cell phenotype. J Exp Med, 1986, 164:351-356.

24. Schmidt RE, Michon JM, Woronicz J, Schlossamn SF, Reinherz EL, and Ritz J. 1987. Enhancement of natural killer function through activation of the T11 E rosette receptor. J Clin Invest 79:305-308.

25. Benjamini E and Leskowitz S. Immunology: A Short Course. Second Edition. New York: Wiley-Liss, 1991, 211-244.

26. Reinherz EL, O'Brien C, Rosenthal P and Schlossman SF. The cellular basis for viral-induced immunodeficiency: Analysis by monoclonal antibodies. J Immunol, 1980, 125:1269-1274.

27. Felsenstein D, Carney WP, Lacoviello VR and Hirsch MS. Phenotypic properties of atypical lymphocytes in cytomegalovirus induced mononucleosis. J Infect Dis, 1985, 152:198-203.

28. Rinaldo CR, Ho M, Hamoudi WH, Gui X and DeBiasio RL. Lymphocyte subsets and natural killer cell responses during cytomegalovirus mononucleosis. Infect Immun, 1983, 40:472-477.

29. Goldstein G, Lifter J and Mittler R. Immunoregulatory changes in human disease detected by monoclonal antibodies to T lymphocytes. In: Monoclonal Antibodies in Clinical Medicine. McMichael AJ and Fabre JW, eds. New York, NY: Academic Press, 1982, pp. 39-70.

30. Schmidt RE: Monoclonal antibodies for diagnosis of Immunodeficiencies. Blut, 1989, 59:200-206.

31. Pedrazzini A, Freedman AS, Andersen J, Heflin L, Anderson K, Takvorian T, Canellos GP, Whitman J, Coral F, Ritz J and Nadler LM. Anti-B cell monoclonal antibody purged autologous bone marrow transplantation for B-cell non-Hodgkin's lymphoma: Phenotypic reconstitution and B-cell function. Blood, 1989, 74:2203-2211.

32. Preijers FWMB, DeWitte T, Wessels JMC, DeGast GC, Van Leeuwen E, Capel PJA and Haanen C. Autologous transplantation of bone marrow purged in vitro with anti-CD7-(WTI-) Ricin A Immunotoxin in T-cell lymphoblastic leukemia and lymphoma. Blood, 1989, 74:1152-1158.

33. Ramos EL, Turka LA, Leggat JE, Wood IG, Milford EL and Carpenter CB. Decrease in phenotypically defined T helper inducer cells (T4+4B4+) and increase in T suppressor effector cells (T8+2H4+) in stable renal allograft recipients. Transplantation, 1989, 47:465-471.

34. Fauci AS: The human deficiency virus: Infectivity and mechanism of pathogenesis. Science, 1988, 239:617-622.

35. Trinchieri G. Biology of natural killer cells. Adv Immunol, 1989, 47:187.

36. Taylor MGJ, Fahey JL, Detels R and Giorgi JV: CD4 percentage, CD4 number and CD4:CD8 ratio in HIV infection: How to choose and how to use. J AIDS, 1989, 2:114-124.

37. Centers for Disease Control and Prevention. Guidelines for performing single-platform absolute CD4+ T-cell determinations with CD45 gating for persons infected with human immunodeficiency virus. MMWR, 2003: 52 (No. RR-2):1-13.

38. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture, Approved Edition (H3). Clinical and Laboratory Standards Institute.

39. Huizinga, T.W., Roos, D., von dem Borne, A.E., "Neutrophil Fc-gamma receptors: A two-way bridge in the immune system", 1990, Mar 15, Blood, 75(6), 1211-1214.

40. Nicholson JKA, Hubbard M and Jones BM. Use of CD45 fluorescence and side scatter characteristics for gating lymphocytes when using the whole blood lysis procedure and flow cytometry. Cytometry (Communications in Clinical Cytometry), 1996, 26:16-21.

41. Cosimi AB, Colvin RB, Burton RC, Rubin RH, Goldstein G, Kung PC, Hansen WP, Delmonico FL and Russell PS. 1981. Use of monoclonal antibodies to T-cell subsets for immunologic monitoring and treatment in recipients of renal allografts. N. Engl. J. Med., 305:308-314.

42. Kaufman A, Herold KC. Anti-CD3 mAbs for treatment of type 1 diabetes. Diabetes Metab Res Rev. 2009 May; 25(4):302-6.

43. Frankel AE, Zuckero SL, Mankin AA, Grable M, Mitchell K, Lee YJ, Neville DM,Woo JH. Anti-CD3 recombinant diphtheria immunotoxin therapy of cutaneous T cell lymphoma. Curr Drug Targets. 2009 Feb; 10(2):104-9.

44. Brunetta E, Hudspeth KL, Mavilio D. Pathologic Natural Killer Cell Subset Redistribution in HIV-1 infection: New Insights in Pathophysiology and Clinical Outcomes. Journal of Leukocyte Biology 2010;88 (Epub ahead of print 07/22/2010).

45. Alter G, Malenfant JM, Delabre RM, Burgett NC, Yu XG, Lichterfeld M, Zaunders J, Altfeld M. Increased Natural Killer Cell Activity in Viremic HIV-1 Infection. The Journal of Immunology 2004(173):5305-5311.

46. CDC: 1997 Revised Guidelines for Performing CD4+ T-Cell Determinations in Persons Infected with Human Immunodeficiency Virus (HIV). MMWR 1997; 46 (RR-2); 1-29.

47. H42-A2 CLSI: Enumeration of Immunologically Defined Cell Populations by Flow Cytometry: Approved Guideline – 2nd Edition 2007.

48. Zloza, et al. Multiple populations of T lymphocytes are distinguished by the level of CD4 and CD8 coexpression and require individual consideration. Journal of Leukocyte Biology 2006:79(4-6).

49. EP06-A CLSI: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline.

50. C28-A23 CLSI: Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline, Third Edition.

51. EP09-A2 CLSI: Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline.

52. EP17-A CLSI: Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Gudieline.

53. Ravetch, J.V., Perussia, B., "Alternative membrane forms of FcγRIII (CD16) on human natural killer cells and neutrophils. Cell type-specific expression of two genes that differ in single nucleotide substitutions." 1989, J. Exp. Med., 170(2), 481-497.

54. Huizinga, T.W., Roos, D., von dem Borne, A.E., "Neutrophil Fc-gamma receptors: A two-way bridge in the immune system", 1990, Mar 15, Blood, 75(6), 1211-1214.

55. Lanier, L.L., Yu, G., Phillips, J.H., "Coassociation of CD3 zeta with a receptor (CD16) for IgG Fc on human natural killer cells", 1989 Dec 14, Nature, 342(6251), 803- 805.

56. Hibbs, M., L., Selvaraj, P., Carpen, O., Springer, T.A., Kuster, H., Jouvin, M. H., Kinet, J.P., "Mechanisms for regulating expression of membrane isoforms of Fc gamma RIII (CD16)", 1989 Dec 22, Science, 246(4937), 1608-1611.

57. Tamm, A., Schmidt, R.E., "The binding epitopes of human CD16 (Fc gamma RIII) monoclonal antibodies: Implications for ligand binding", 1996 Aug 15, J. Immunol., 157(4), 1576-1581.

58. Tamm, A., Bassmann, Schmidt, R.E., "Natural killer cell structural studies: Localization of the epitopes of human CD16 (FcγRIII) monoclonal antibodies on the molecular model of CD16", 1997, Leucocyte Typing VI, White Cell Differentiation Antigens, Kishimoto, T., et al, Eds., Garland Publishing, Inc., 324-326.

59. Ritz, J., Trinchieri, G., Lanier, L.L., "NKcell antigens: section report", 1995, Leucocyte Typing V, White Cell Differentiation Antigens. Schlossman, S.F., et al., Eds., Oxford University Press, 1367-1372.

60. Tamm, A., Kister, A., Nolte, K.U., Gessner, J.E., and Schmidt, R.E. "The IgG binding site of human Fc gamma RIIIB receptor involves CC' and FG loops of the membrane-proximal domain." 1996 Feb 16, J Biol Chem, 271(7), 3659-66.

61. Denny TN, Gelman R, Bergeron M, Landay A et al. A North American multilaboratory study of CD4 counts using flow cytometric panLeukogating (PLG): a NIAID-DAIDS Immunology Quality Assessment Program Study. Cytometry B Clin Cytom. 2008;74 Suppl 1:S52-64.

62. Glencross D, Scott LE, Jani IV, Barnett D CD45-assisted PanLeucogating for accurate, cost-effective dual-platform CD4+ T-cell enumeration. Cytometry. 2002 Apr;15;50(2):69-77.

63. Reimann KA, O'Gorman MR, Spritzler J, et al. Multisite comparison of CD4 and CD8 T-lymphocyte counting by single- versus multiple-platform methodologies: evaluation of Beckman Coulter Flow-Count Fluorospheres and the tetraONE system. The NIAID DAIDS New Technologies Evaluation Group. Clin Diagn Lab Immunol. 2000 May;7(3):344-51.

64. Suades R, Padró T, Alonso R, et al. Circulating CD45+/CD3+ lymphocyte-derived microparticles map lipid-rich atherosclerotic plaques in familial hypercholesterolaemia patients. Thromb Haemost. 2014 Jan;111(1):111-21

of 40

5 of 5

PRODUCT AVAILABILITY AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5

B23533 - 50 tests (0.9 mL)

AQUIOS Tetra-2+ Panel CD45-FITC/(CD56+CD16)-RD1/CD19-ECD/CD3-PC5

B23534 - 50 tests (0.9 mL)

TRADEMARKS AQUIOS, Beckman Coulter and the stylized logo are trademarks of Beckman Coulter, Inc. Beckman Coulter and the stylized logo are registered in the USPTO. Texas Red-X is a trademark of Molecular Probes, Inc. For additional information, or if damaged product is received, call Beckman Coulter Customer Service at 800-526-7694 (USA or Canada) or contact your local Beckman Coulter Representative.

Beckman Coulter Ireland Inc. Lismeehan O’Callaghan’s Mills Co.Clare, Ireland 353 (0)65 683 1100 www.beckmancoulter.com

© 2015 Beckman Coulter, Inc. All Rights Reserved.

Revision History Revision AA, 12/2013

� Initial Release

Revision AB, 12/2014 � Statement of Warnings

� Storage Conditions and Stability

Revision AC, 4/2015 � Intended Use

� Summary and Explanation � Clinical Relevance

� Storage Conditions and Stability � Limitations � Specificity

� References

of 40

1 of 2

For In Vitro Diagnostic Use

Rx Only

INTENDED USE AQUIOS IMMUNO-TROL Low Cells is an assayed, lysable whole blood quality control product for immunophenotyping analysis using monoclonal antibody reagents and flow cytometry. It provides a positive cell control that is processed in the same manner as a whole blood sample. This allows verification of instrument and reagent performance. It also verifies the methods used for staining targeted cells, the lysing of erythrocytes, and the analysis of samples by the AQUIOS CL Flow Cytometer.

SUMMARY AND EXPLANATION Immunophenotyping analysis using flow cytometry involves the identification and enumeration of targeted cells in whole blood samples. Whole blood samples are stained with monoclonal antibodies and erythrocytes are lysed prior to flow cytometric analysis. A Positive cell control is required to verify reagent performance, sample preparation methods, and staining procedures.1,2 A positive cell control should mimic a representative whole blood sample in terms of monoclonal antibody performance, erythrocyte lysing, and flow cytometric analysis. AQUIOS IMMUNO-TROL Low Cells is a liquid preparation of stabilized human erythrocytes and leukocytes (lymphocytes, monocytes, and granulocytes) that have lysing, light scatter, antigen expression, and antibody staining properties representative of those found in human normal whole blood.

PRINCIPLES OF TEST AQUIOS IMMUNO-TROL Low Cells consists of two major cell components: leukocytes and erythrocytes. The leukocytes function as the positive cell component of AQUIOS IMMUNO-TROL Low Cells and have surface antigens present on the targeted cells that bind to the antibody component of the monoclonal antibody reagent. The erythrocytes function as the lysable component of AQUIOS IMMUNO-TROL Low Cells. AQUIOS IMMUNO-TROL Low Cells samples are first stained with the monoclonal antibody reagents and then lysed to remove erythrocytes. Flow cytometric analysis of the stained and lysed AQUIOS IMMUNO-TROL Low Cells samples determines the percentage and absolute count of the targeted cells. Expected Results are determined on validated and standardized AQUIOS Systems.

REAGENT AQUIOS IMMUNO-TROL Low Cells is a liquid preparation of stabilized human erythrocytes and leukocytes in a stabilizing solution containing BSA.

When using Tetra-1 Panel or Tetra-2+ Panel tests, there are 15 tests per tube. When using the Tetra Combo, there are 10 tests per tube. Each cap should have no more than 15 piercings total.


Neomycin Sulfate <0.1%

Penicillin-G, sodium salt <0.1%



1. This reagent contains <0.1% sodium azide. Sodium azide under acid conditions yields hydrazoic acid, an extremely toxic compound. Azide compounds should be flushed with running water while being discarded. These precautions are recommended to avoid deposits in metal piping in which explosive conditions can develop. If skin or eye contact occurs, wash excessively with water.

2. POTENTIAL BIOHAZARDOUS MATERIAL Each donor unit used in the preparation of this material was tested by an FDA-approved method for the presence of the antibodies to Human Immunodeficiency Virus (HIV-1, HIV-2) and Hepatitis C Virus (HCV) as well as for Hepatitis B surface antigen (HBsAg) and found to be negative (were not repeatedly reactive). Because no test method can offer complete assurance that HIV, HCV, Hepatitis B Virus (HBV), or other infectious agents are absent, this reagent should be handled at the Biosafety Level 2 as recommended for any potentially infectious human serum or blood specimen in the Centers for Disease Control/National Institutes of Health manual "Biosafety in Microbiological and Biomedical Laboratories," 1988.

3. Never pipet by mouth and avoid contact of samples with skin and mucous membranes.

4. Do not freeze reagents.

5. Avoid microbial contamination of reagents or erroneous results may occur.

6. Use Good Laboratory Practices (GLP) when handling this reagent.

STORAGE AND STABILITY Do not use reagent beyond the expiration date printed on the vial label. Store upright. Unopened reagent is stable to the expiration date on the vial label when stored at 2-8°C. Opened reagent is stable for 90 days after opening when stored at 2-8°C. Do not freeze.

EVIDENCE OF DETERIORATION Inability to obtain expected results or a shift in light scatter or fluorescence properties may indicate product deterioration. Instrument standardization, sample preparation, and reagents, antibody staining and antibody performance should also be investigated. Hemolysis may indicate improper storage conditions that may affect product performance in terms of lysability.

PROCEDURE

1. For every new lot of controls, select the barcode button on the AQUIOS Software Main Screen and scan the assay sheet barcode to load assay values.

NOTE: In the event the barcode fails, refer to the TO ENTER ASSAY VALUES MANUALLY section.

2. Bring the control reagent and antibodies to room temperature.

3. Gently roll/invert the control reagent to mix the product in a manner similar to the handling of a whole blood specimen.

4. Verify that a QC request has been created.

5. Select “QC” in the “Test Request”.

6. LOAD Control(s), with cap(s) on, on the AQUIOS system.

� For autoloader, insert Control tube(s) into the instrument cassette and place the cassette on the system.

� For single tube loader, mix the Control immediately before placing the tube in the tube sampling area. Scan the tube and place on the system.

7. GO is automatically initiated when Control(s) are placed on the system.

� For autoloader, the system runs the Control(s) when the cassette is loaded.

� For single tube loader, the system runs the Control(s) when the door is closed.

8. Allow system to prepare controls and to analyze.

9. When the system is done with the tube, store Control(s) upright in refrigerator.

TO ENTER ASSAY VALUES MANUALLY:

1. Select Setup.

2. Select Reagent Info.

3. Select Edit.

4. Select IMMUNO-TROL Cells or IMMUNO-TROL Low Cells from the Reagent/Sample drop-down list.

5. Select the Lot Number from the Lot Number drop-down list or click on + to add a new Lot Number.

6. Enter the Expiration Date.

7. Enter the values from the Assay Sheet into the table Min and Max columns. NOTE: Percentages must be entered as decimals.

8. Select Save. Refer to the Instructions For Use (IFU) PN B21896 for detailed information on logging in control values.

EXPECTED RESULTS The mean values are determined for each antigen using aN AQUIOS flow cytometer. Under these conditions, 95% of the recovered values should fall within the stated expected range. Each laboratory should establish its own mean and expected range for each antigen analyzed. Participation in Beckman Coulter's Interlaboratory Quality Assurance Program (IQAP) provides a comparison to peer laboratories and augments a laboratory’s quality assurance program. IMPORTANT: Do NOT move the regions or modify compensation for control runs. Data presented is representative of expected performance. Figure 1 – CD45-FITC vs Side Scatter white blood cell populations are defined. A first Gating region identified lymphocytes.

AQUIOS IMMUNO-TROL

Low Cells

B25700

PN B29141-AB

of 40

2 of 2

Figure 2 - EV/SS (Electronic Volume vs Side Scatter) dot plot is created to optimize the population of interest and define the second lymph gate.

NOTE: This plot is viewable from the Detail Report screen.

Figure 3. CD3+ Cells: CD3+ALL (CD3-PC5) is defined by gating Lymphs All Region previously defined by CD45 Side Scatter (Figure 1) or CD45-EV (Figure 2) CD45-EV one or two.

Figure 4 – Plot of CD4-RD1 vs CD8-ECD derived from CD3+ All

Figure 5 - Plot of CD56/16-RD1 vs. CD19-ECD from CD3-

LIMITATIONS

1. Erroneous results may occur if the flow cytometer is not properly aligned or standardized for fluorescence or if the cell populations are improperly gated.

2. Results determined using flow cytometers, lysing systems, or antibodies that are different from those used to determine the expected results, may not fall within the expected range.

REFERENCES

1. National Committee for Clinical Laboratory Standards. Clinical Applications of Flow Cytometry: Quality Assurance and Immunophenotyping of Lymphocytes; Approved Guideline. 1998. NCCLS document H42-A.

2. 1997 Revised Guidelines for the Performance of CD4+ T-Cell Determination in Persons with Human Immunodeficiency Virus (HIV) Infection. MMWR, January 10, 1997, p.11.

PRODUCT AVAILABILITY AQUIOS IMMUNO-TROL Low Cells

B25700 (2 x 3 mL vials)

TRADEMARKS AQUIOS, Beckman Coulter and the stylized logo are trademarks of Beckman Coulter, Inc. Beckman Coulter and the stylized logo are registered in the USPTO. For additional information, or if damaged product is received, call Beckman Coulter Customer Service at 800-526-7694 (USA or Canada) or contact your local Beckman Coulter Representative.

Beckman Coulter Ireland Inc.Lismeehan O'Callaghan's Mills Co. Clare, Ireland 353 (0)65 683 1100 www.beckmancoulter.com


Revision History

Revision AA, 12/2013 � Initial Release


of 40

1 of 2

For In Vitro Diagnostic Use

Rx Only

INTENDED USE AQUIOS IMMUNO-TROL Cells is an assayed, lysable whole blood quality control product for immunophenotyping analysis using monoclonal antibody reagents and flow cytometry. It provides a positive cell control that is processed in the same manner as a whole blood sample. This allows verification of instrument and reagent performance. It also verifies the methods used for staining targeted cells, the lysing of erythrocytes, and the analysis of samples by the AQUIOS CL Flow Cytometer.

SUMMARY AND EXPLANATION Immunophenotyping analysis using flow cytometry involves the identification and enumeration of targeted cells in whole blood samples. Whole blood samples are stained with monoclonal antibodies and erythrocytes are lysed prior to flow cytometric analysis. A positive cell control is required to verify reagent performance, sample preparation methods and staining procedures.1,2 A positive cell control should mimic a representative whole blood sample in terms of monoclonal antibody performance, erythrocyte lysing, and flow cytometric analysis. AQUIOS IMMUNO-TROL Cells is a liquid preparation of stabilized human erythrocytes and leukocytes (lymphocytes, monocytes, and granulocytes) that have lysing, light scatter, antigen expression, and antibody staining properties representative of those found in human normal whole blood.

PRINCIPLES OF TEST AQUIOS IMMUNO-TROL Cells consists of two major cell components: leukocytes and erythrocytes. The leukocytes function as the positive cell component of AQUIOS IMMUNO-TROL Cells and have surface antigens present on the targeted cells that bind to the antibody component of the monoclonal antibody reagent. The erythrocytes function as the lysable component of AQUIOS IMMUNO-TROL Cells. AQUIOS IMMUNO-TROL samples are first stained with the monoclonal antibody reagents and then lysed to remove erythrocytes. Flow cytometric analysis of the stained and lysed AQUIOS IMMUNO-TROL samples determines the percentage and absolute count of the targeted cells. Expected Results are determined on validated and standardized AQUIOS Systems.

REAGENT AQUIOS IMMUNO-TROL Cells is a liquid preparation of stabilized human erythrocytes and leukocytes in a stabilizing solution containing BSA.

When using Tetra-1 Panel or Tetra-2+ Panel tests, there are 15 tests per tube. When using the Tetra Combo, there are 10 tests per tube. Each cap should have no more than 15 piercings total.


Neomycin Sulfate <0.1%

Penicillin-G, sodium salt <0.1%



1. This reagent contains <0.1% sodium azide. Sodium azide under acid conditions yields hydrazoic acid, an extremely toxic compound. Azide compounds should be flushed with running water while being discarded. These precautions are recommended to avoid deposits in metal piping in which explosive conditions can develop. If skin or eye contact occurs, wash excessively with water.

2. POTENTIAL BIOHAZARDOUS MATERIAL Each donor unit used in the preparation of this material was tested by an FDA-approved method for the presence of the antibodies to Human Immunodeficiency Virus (HIV-1, HIV-2) and Hepatitis C Virus (HCV) as well as for Hepatitis B surface antigen (HBsAg) and found to be negative (were not repeatedly reactive). Because no test method can offer complete assurance that HIV, HCV, Hepatitis B Virus (HBV), or other infectious agents are absent, this reagent should be handled at the Biosafety Level 2 as recommended for any potentially infectious human serum or blood specimen in the Centers for Disease Control/National Institutes of Health manual "Biosafety in Microbiological and Biomedical Laboratories," 1988.

3. Never pipet by mouth and avoid contact of samples with skin and mucous membranes.

4. Do not freeze reagents.

5. Avoid microbial contamination of reagents or erroneous results may occur.

6. Use Good Laboratory Practices (GLP) when handling this reagent.

STORAGE AND STABILITY Do not use reagent beyond the expiration date printed on the vial label. Store upright. Unopened reagent is stable to the expiration date on the vial label when stored at 2-8°C. Opened reagent is stable for 90 days after opening when stored at 2-8°C. Do not freeze.

EVIDENCE OF DETERIORATION Inability to obtain expected results or a shift in light scatter or fluorescence properties may indicate product deterioration. Instrument standardization, sample preparation, and reagents, antibody staining and antibody performance should also be investigated. Hemolysis may indicate improper storage conditions that may affect product performance in terms of lysability.

PROCEDURE

1. For every new lot of controls, select the barcode button on the AQUIOS Software Main Screen and scan the assay sheet barcode to load assay values.

NOTE: In the event the barcode fails, refer to the TO ENTER ASSAY VALUES MANUALLY section.

2. Bring the control reagent and antibodies to room temperature.

3. Gently roll/invert the control reagent to mix the product in a manner similar to the handling of a whole blood specimen.

4. Verify that a QC request has been created.

5. Select “QC” in the “Test Request”.

6. LOAD Control(s), with cap(s) on, on the AQUIOS system.

� For autoloader, insert Control tube(s) into the instrument cassette and place the cassette on the system.

� For single tube loader, mix the Control immediately before placing the tube in the tube sampling area. Scan the tube and place on the system.

7. GO is automatically initiated when Control(s) are placed on the system.

� For autoloader, the system runs the Control(s) when the cassette is loaded.

� For single tube loader, the system runs the Control(s) when the door is closed.

8. Allow system to prepare controls and to analyze.

9. When the system is done with the tube, store Control(s) upright in refrigerator.

TO ENTER ASSAY VALUES MANUALLY:

1. Select Setup.

2. Select Reagent Info.

3. Select Edit.

4. Select IMMUNO-TROL Cells or IMMUNO-TROL Low Cells from the Reagent/Sample drop-down list.

5. Select the Lot Number from the Lot Number drop-down list or click on + to add a new Lot Number.

6. Enter the Expiration Date.

7. Enter the values from the Assay Sheet into the table Min and Max columns. NOTE: Percentages must be entered as decimals.

8. Select Save. Refer to the Instructions For Use (IFU) PN B21896 for detailed information on logging in control values.

EXPECTED RESULTS The mean values are determined for each antigen using an AQUIOS flow cytometer. Under these conditions, 95% of the recovered values should fall within the stated expected range. Each laboratory should establish its own mean and expected range for each antigen analyzed. Participation in Beckman Coulter's Interlaboratory Quality Assurance Program (IQAP) provides a comparison to peer laboratories and augments a laboratory's quality assurance program. IMPORTANT: Do NOT move the regions or modify compensation for control runs.

Data presented is representative of expected performance. Figure 1. CD45-FITC vs Side Scatter white blood cell populations are defined. A first Gating region identified lymphocytes

AQUIOS IMMUNO-TROL

Cells

B23535

PN B29142-AB

of 40

2 of 2

Figure 2 - EV/SS (Electronic Volume vs Side Scatter) dot plot is created to optimize the population of interest and define the second lymph gate.

NOTE: This plot is viewable from the Detail Report screen.

Figure 3. CD3+ Cells: CD3+ALL (CD3-PC5) is defined by gating Lymphs All Region previously defined by CD45 Side Scatter (Figure 1) or CD45-EV (Figure 2) CD45-EV one or two.

Figure 4. Plot of CD4-RD1 vs CD8-ECD derived from CD3+ All

Figure 5. Plot of CD56/16-RD1 vs. CD19-ECD from CD3-

LIMITATIONS

1. Erroneous results may occur if the flow cytometer is not properly aligned or standardized for fluorescence or if the cell populations are improperly gated.

2. Results determined using flow cytometers, lysing systems, or antibodies that are different from those used to determine the expected results, may not fall within the expected range.

REFERENCES

1. National Committee for Clinical Laboratory Standards. Clinical Applications of Flow Cytometry: Quality Assurance and Immunophenotyping of Lymphocytes; Approved Guideline. 1998. NCCLS document H42-A.

2. 1997 Revised Guidelines for the Performance of CD4+ T-Cell Determination in Persons with Human Immunodeficiency Virus (HIV) Infection. MMWR, January 10, 1997, p.11.

PRODUCT AVAILABILITY AQUIOS IMMUNO-TROL Cells

B23535 (2 x 3 mL vials)

TRADEMARKS AQUIOS, Beckman Coulter and the stylized logo are trademarks of Beckman Coulter, Inc. Beckman Coulter and the stylized logo are registered in the USPTO. For additional information, or if damaged product is received, call Beckman Coulter Customer Service at 800-526-7694 (USA or Canada) or contact your local Beckman Coulter Representative.

Beckman Coulter Ireland Inc.Lismeehan O'Callaghan's Mills Co. Clare, Ireland 353 (0)65 683 1100 www.beckmancoulter.com


Revision History

Revision AA, 12/2013 � Initial Release


of 40

PQDx 0156 053 00 WHO PQDx PR November/2015, … 0156‐053‐00 WHO PQDx PR November/2015, version...

Documents

Transcript of PQDx 0156 053 00 WHO PQDx PR November/2015, … 0156‐053‐00 WHO PQDx PR November/2015, version...