ARCHITECT Total T3 Assay Troubleshooting...

ABBOTT LABORATORIES Part Number 201422-102Abbott Park, IL 60064 © 2005, 2010Architect_TT3.fm, Revision 2 CONFIDENTIAL

ARCHITECT Total T3 AssayTroubleshooting Guide

2-Step Assay Using CMIA TechnologyWW (except U.S.) Use U.S. Only Use WW Use

• Reagents / Calibrators / ControlsList Numbers Control Kit MasterCheckReagent Size Codes Calibrator Kit Multiconstituent Control Kit(s)Reagent Kit Data Reduction

• Sample InformationDiagnostic Purpose Minimum Sample VolumeSample Type Panel Testing

• Clinical ResultsExpected Values SpecificityPrecision InterferenceRecovery CorrelationSensitivity Method Comparison

• Assay File / Protocol InformationAssay Parameters Error Code / Observed ProblemDilution

• KeywordsAlternate Result Unit Conversion Frozen Specimen Storage Reference Assay SequenceAnalytical Sensitivity Protocol Functional Sensitivity Reference MethodAssay CD-ROM Intended Use Reportable Range VerificationAssay Sequence Least Squares Correlation Plot(s) “Same As” ListCalibration Validity Specifications Levey-Jennings Setting Control RangesCompetitor Assay Info Matrix Effects Shipping ConditionsContents Microbial Contamination Specimen HandlingCurve Shape Onboard Stability StandardizationCurve Stability Panel Requirements StorageEvaporation Passing-Bablok Correlation Plot(s) Summary and Explanation of Test

Reagent Appearance Westgard Rules

• Package Insert Bibliography

• Revision HistoryFOR INTERNAL USE

July 2010

ARCHITECT Total T3 AssayInformation in red italicized text comes from applicable Package Inserts and/or the assay Launch Binder.

WW (except U.S.) Use U.S. Only Use WW Use

Reagents / Calibrators / Controls

Category Description KeywordList Numbers Reagents: 7K64

Controls: NACalibrators: 7K64-01

Immunoassay MCC* (liquid): 6E20-10Manual Diluent: 7K64-50

*Multi-constituent Control

• Assay CD-ROM• Shipping

Conditions• Storage

Reagent Size Codes

Reagent Kit • Assay Sequence• Onboard

Stability• Reagent

Appearance• Reference Assay

Sequence• Shipping

Conditions• Storage

Control Kit(SCC*)

ARCHITECT Total T3 controls are not available.

Calibrator Kit Standardization Information: The calibrators are manufactured using L-Triiodothyronine, sodium salt (USP Grade) and signal matched to Abbott internal reference standards.

• Alternate Result Unit Conversion

• Contents• Curve Shape• Curve Stability• “Same As” List• Shipping

Conditions• Standardization• Storage

Size Code Description

-20 4 X 100 Test Kit



Minimum Volume(100/500 Test) Band Color Contents

5.9 mL /26.3 mL Yellow

T3 acridinium-labeled Conjugate in citrate buffer with NaCl and Triton X-100 stabilizers.Minimum concentration: 0.33 ng/mL. Preservative: antimicrobial agent.

6.6 mL /27.0 mL PinkAnti-T3 (sheep) coated Microparticles in MES buffer with sheep IgG stabilizers. Preservative: antimicrobial agent.

LevelMinimum Vol.

(mL) Target

(ng/mL)Target

(nmol/L)

Cal 1 4 0.5 0.77

Cal 2 4 8.0 12.3

Part Number 201422-102Architect_TT3.fm, Revision 2 1 of 26 CONFIDENTIAL



Data Reduction • Calibration Method: 2-pt calibration• Calibration Adjustment Method: 4 PLC, Y-weighted Linear

Transformation

** NOTE: The above information SHOULD NOT be used to judge the validity of a customer’s calibration curve on this reagent. The Average RLU and Estimated RLU range are examples based on internal studies.

• Calibration Validity Specifications

• Curve Shape• Curve Stability

MasterCheck ARCHITECT MasterCheck was used for the verification of calibration/reportable range and sensitivity of the assay based on CLIA regulations. This product is no longer manufactured for ARCHITECT assays. It is recommended that customers use other suitable samples for verification procedures.

• Analytical Sensitivity Protocol

• Reportable Range Verification

Multiconstituent Control Kit(s)

NOTE: See lot specific MCC Data Sheet for initial control ranges.

• Contents• Levey-Jennings• Matrix Effects• Setting Control

Ranges• Shipping

Conditions• Storage• Westgard Rules

Reagents / Calibrators / Controls

Category Description Keyword

CalibratorConcentration

(ng/mL)AverageRLU’s Estimated RLU Range**

A 0.0 235650 179752 - 305968

B/1 0.5 183257 130812 - 243167

C 1.0 133535 88543 - 184648

D 2.0 62736 35456 - 96889

E 4.0 20583 14646 - 28806

F/2 8.0 13220 10530 - 17525

Name List LevelsMinimum Vol.

(mL)

Immunoassay - MCC (Liquid) 06E20-10 1, 2, 3 5




Sample Information

Category Description KeywordDiagnosticPurpose

3,5,3’ Triiodothyronine (T3) is a thyroid hormone with a molecular weight of 651 daltons1 and a half-life in serum of 1.5 days.2 T3 circulates in the blood as an equilibrium mixture of free and protein bound hormone.3 T3 is bound to thyroxine binding globulin (TBG), prealbumin, and albumin. The actual distribution of T3 among these binding proteins is controversial as estimates range from 38-80% for TBG, 9-27% for prealbumin, and 11-35% for albumin.4 The binding of these proteins is such that only 0.2-0.4% of the total T3 is present in solution as unbound or free T3.5 This free fraction represents the physiologically active thyroid hormone.3

It has become apparent in recent years that T3 plays an important role in the maintenance of the euthyroid state. Serum T3 measurements can be a valuable component of a thyroid screening panel in diagnosing certain disorders of thyroid function as well as conditions caused by iodine deficiency. Clinically, measurements of serum T3 concentration are especially valuable in diagnosing hyperthyroidism and in following the course of therapy for this disorder.2,6,7 Under conditions of strong thyroid stimulation, the T3 measurement provides a good estimation of thyroid reserve.2 Recognition of a thyroid dysfunction called T3-thyrotoxicosis, associated with an increased serum T3 level but normal thyroxine (T4), free T4, and in vitro Uptake results have further highlighted the importance of serum T3 measurements.2,8-11 Dietary iodine deficiency results in inadequate production of thyroid hormones despite the presence of normal thyroid tissue. In these cases, the serum T4 concentration is often low while the thyroid stimulating hormone (TSH) concentration is elevated. Elevated TSH associated with low T4 is normally indicative of hypothyroidism. However, in iodine deficiency, these results together with normal or slightly elevated serum T3 are indicative of euthyroid status in most individuals.12

• Intended Use• Package Insert

Bibliography• Summary and

Explanation of Test




Sample Type • Frozen Specimen Storage

• Shipping Conditions

• Specimen Handling

Minimum Sample Volume

Calibrators: 4 dropsControls: N/A

Sample - Single Test: 150 µLSample - Each Additional Test: 25 µL

Priority Sample: 25 µL for each replicate plus50 µL for dead volume

Replicate Limitations: No more than 10 replicates maybe sampled from the same samplecup in order to avoid dilutioneffects.

• Evaporation

Panel Testing Assays that would be tested as a panel:• Free T4• Total T4• Free T3• Total T3• TSH

• Panel Requirements

Sample Information


Specimen Type(s):

• Human Serum and Plasma (including serum collected in serum separator tubes)

• Plasma collected in sodium heparin, lithium heparin, or potassium EDTA anticoagulant tubes

Note: Other specimen types have not been validated for use.

Specimen Storage:

Serum or PlasmaIf testing will be delayed more than 24 hours, remove serum or plasma from the clot, serum separator, or red blood cells.

Type Temperature TimeSerum 2-8°C 6 days

-10°C 6 days

Plasma 2-8°C 6 days

-10°C 6 days




Clinical Results

Category Description KeywordExpected Values A normal range of 0.58 ng/mL (0.89 nmol/L) to 1.59 ng/mL (2.44

nmol/L) (central 95% interval) was obtained by testing serum specimens from 438 individuals determined as normal by AxSYM Ultrasensitive hTSH II and AxSYM Free T4 assays. It is recommended that each laboratory establish its own normal range, which may be unique to the population it serves depending upon geographical, patient, dietary, or environmental factors.

Precision The ARCHITECT Total T3 assay is designed to have a precision of < 10% (total CV). A study based on guidance from (NCCLS) document EP5-A22 was performed for the ARCHITECT Total T3 assay. A three member processed human serum based panel was assayed, using two lots of reagents, in replicates of two at two separate times per day for 20 testing days. Data from this study are summarized in the following table.*

* Representative data; results in individual laboratories may vary from these data.


Panel Member

ReagentLot

Mean Conc. Value Within Run Total

Instrument n (pg/mL) SD %CV SD %CV1 1 1 80 0.75 0.021 2.7 0.027 3.61 1 2 80 0.73 0.023 3.1 0.030 4.11 2 1 80 0.79 0.036 4.6 0.043 5.41 2 2 80 0.81 0.047 5.8 0.057 7.02 1 1 80 1.50 0.029 1.9 0.110 7.32 1 2 80 1.49 0.040 2.7 0.053 3.62 2 1 80 1.53 0.031 2.0 0.035 2.32 2 2 80 1.54 0.040 2.6 0.049 3.23 1 1 80 3.27 0.062 1.9 0.128 3.93 1 2 80 3.29 0.107 3.3 0.140 4.23 2 1 80 3.55 0.054 1.5 0.071 2.03 2 2 80 3.54 0.066 1.9 0.077 2.2




Recovery The ARCHITECT Total T3 assay is designed to have a mean recovery of 100 +/- 10% when analyzing samples spiked with known amounts of T3. T3 was added to nine normal human serum samples. The concentration of T3 was determined using the ARCHITECT Total T3 assay and the resulting percent recovery was calculated.*

Average Recovery: 98.6%* Representative data; results in individual laboratories may vary from these data.

Sensitivity The ARCHITECT Total T3 assay is designed to have an analytical sensitivity of < 0.25 ng/mL.Analytical sensitivity is defined as the concentration calculated as the mean plus two standard deviations of replicates of the ARCHITECT Total T3 MasterCheck Level 0 (0.0 ng/mL). The analytical sensitivity (low-linearity) is defined in the ARCHITECT Total T3 assay parameters as 0.25 ng/mL.

MasterCheck are no longer manufactured for the ARCHITECT Total T3 assay.

• Analytical Sensitivity Protocol

• Functional Sensitivity

Specificity The ARCHITECT Total T3 assay is designed to have a mean analytical specificity of < 0.1% cross reactivity with thyroxine (T4) at a concentration of 1,100 ng/mL.

Interference The ARCHITECT Total T3 assay is designed to have a mean potential interference from hemoglobin, bilirubin, triglycerides, and protein of < 10% at the levels indicated below.

• Matrix Effects• Microbial

Contamination

Clinical Results


Sample

Endogenous Total T3

Conc.(ng/mL)

T3Conc.

(ng/mL)

ObservedTotal T3

Conc.(ng/mL)

%Recovery

1 2.01 0.77 2.74 94.8

2 0.97 0.78 1.64 85.9

3 1.13 0.79 1.95 103.8

4 0.99 1.54 2.43 93.5

5 0.88 1.53 2.41 100.0

6 0.90 1.54 2.54 106.5

7 1.07 3.03 4.28 105.9

8 1.23 3.04 4.21 98.0

9 0.90 3.03 3.89 98.7

Interfering Substance Concentration InterferenceHemoglobin <500 mg/dL <10%

Bilirubin <20 mg/dL <10%Triglycerides <2000 mg/dL <10%

Protein <12 g/dL <10%




Correlation The ARCHITECT Total T3 assay is designed to have a slope of 1.00 + 0.20 and a correlation coefficient (r) of > 0.90 when compared to the AxSYM Total T3 assay.A study was performed where specimens were tested using the ARCHITECT Total T3 assay and AxSYM Total T3 assay. Data from this study were analyzed using least squares and Passing-Bablok23 regression methods and are summarized in the following table.*

Abbott ARCHITECT Total T3 vs. Abbott AxSYM Total T3

In this evaluation, serum specimens tested ranged from 0.25 ng/mL (0.38 nmol/L) to 5.83 ng/mL (8.95 nmol/L) with the ARCHITECT Total T3 assay and from 0.34 ng/mL (0.52 nmol/L) to 5.19 ng/mL (7.97 nmol/L) with the AxSYM Total T3 assay.* Representative data; variables such as differences in sampling

size and sample population may impact the correlation of the assay; therefore, results in individual laboratories may vary from these data.

** A linear regression method with no special assumptions regarding the distribution of the samples and measurement errors.19

• Competitor Assay Info

• Least Squares Correlation Plot(s)


• Passing-Bablok Correlation Plot(s)

• Reference Assay Sequence

• Reference Method

Clinical Results


Method n

Correlation Coefficient

(r) Slope Intercept

Least SquaresLinear Regression 1440 0.964 1.04 0.02

Passing BablokLinear Regression** 1440 0.964 1.13 -0.08




Method Comparison

aReference: ARCHITECT System Operations ManualbReference: AxSYM System Operations Manual

• Competitor Assay Info

• Reference Assay Sequence

Clinical Results


Assay Parameters ARCHITECT AxSYM

Assay PrincipleChemiluminescent

Microparticle Immunoassay (CMIA)

Microparticle Enzyme Immunoassay (MEIA)

Throughput 29 min. (time to 1st result)200 tests/houra

26 min. (time to 1st result in batch mode)

46 tests/hr mixed66 tests/hr batch

Analyte Measured Triiodothyronine Triiodothyronine

Sample Size 25-150 µL 115-150 µL

Calibrator Targets 0.5 and 8.0 ng/mL 0, 0.5, 1, 2, 4, and 8 ng/mL

Control Targets N/ALow = 0.7 ng/mL

Medium = 1.5 ng/mLHigh = 3.7 ng/mL

Standardization USP Reference Standard L-Triiodothyronine

USP Reference Standard L-Triiodothyronine

Specimen Type(s) Human Serum or Plasma Human Serum or Plasma

Assay Range (Reportable Range) 0.25-8.0 ng/mL 0.30-8 ng/mL

Expected Values 0.58-1.59 ng/mL 0.79-1.49 ng/mL

Precision<10% (total CV)

Precision

<8% (total CV) for concentrations in the high

control range<10% (total CV) for

concentrations in the medium control range

<16% (total CV) for concentrations in the low

control range

Sensitivity <0.25 ng/mL <0.30 ng/mL

Assay Temperature 37.0 +/- 0.6°Ca 34.0 +/-0.5°Cb




Assay File / Protocol Information


AssayParameters

The default values for the assay parameters used in the ARCHITECT Total T3 assay are contained in the categories listed below. Assay parameters that can be edited contain a (>). In order to obtain values for the parameters with an (*), review the category specific details for assay parameters dialog windows. For detailed information on viewing and editing assay parameters, refer to the ARCHITECT i System Operations Manual, Section 2.For a detailed description of system procedures, refer to the ARCHITECT i System Operations Manual.

• Alternate Result Unit Conversion

General Parameters

Name: > _TT3

Assay Type: Two Step 18 4

Assay Number: 671

Pretreatment option: None

Assay Version: 7

Assay Availability: *

Cal Version: 2

Assay Status:> *

Run Controls for reagent by:>

Calibration Parameters

Type: Adjust

Calibration Method: 4PLC X Transform

Adjustment Method: Linear Transformation

Replicates: > 2

Cal A: 0.0

Cal B: 0.5

Cal C: 1.0

Cal D: 2.0

Cal E: 4.0

Cal F: 8.0

Cal 1: 0.5

Cal 2: 8.0

Dilution Parameters

Manual Dilution: On

Default Dilution: Undiluted

Dilution Range Low High 0.25 8.0

Results Parameters

Linearity Range: 0.25 - 8.0

Normal Range: >

Extreme Range: >

Results Units

Result concentration units: ng/mL

Result decimal places: >2




Dilution Specimens with a Total T3 value exceeding 8.00 ng/mL (12.29 nmol/L), are flagged with the code “>8.00” and may be diluted with the Manual Dilution Procedure.• Manual Dilutions should be performed as follows:• The suggested dilution for Total T3 is 1:2. It is recommended

dilutions not exceed 1:2.• Specimen and serial dilutions greater than 1:2 have not been

validated for use by the ARCHITECT Total T3 assay.• For a 1:2 dilution, add a minimum of 75 µL of the patient

specimen to 75 µL of ARCHITECT Total T3 Manual Diluent.• To avoid contamination of Manual Diluent, dispense several

drops of Manual Diluent into a clean test tube prior to pipetting.• The operator must enter the dilution factor (2) in the patient or

control order screen. The system will use this dilution factor to automatically calculate the concentration of the sample before dilution. This will be the reported result. The dilution should be performed so that the reported result reads greater than 1.0 ng/mL (1.536 nmol/L).

• If the operator does not enter the dilution factor, the reported result will be that of the diluted sample. This result (before dilution factor is applied) should be greater than 0.5 ng/mL (0.77 nmol/L).

• For detailed information on ordering dilutions, refer to the ARCHITECT i System Operations Manual, Section 5.

Error Code / Observed Problem

The ARCHITECT Total T3 assay has not experienced any assay-specific errors.

Assay File / Protocol Information





Keywords InformationAlternate Result Unit Conversion

• Default result unit: ng/mL• Alternate Result Unit: nmol/L• Conversion Factor: 1.536• Conversion Formula:(Concentration in ng/mL) x (1.536) = nmol/L• Alternate Result Unit: ng/dL• Conversion Factor: 100• Conversion Formula: (Concentration in ng/mL) x (100.0) =ng/dL*

* i System Assay CD-ROM version 6.0 and higher will be required to install this alternate result unit (ng/dL).

Analytical Sensitivity Protocol

• Refer to the ARCHITECT i System Operations Manual, Appendix B• Group Number: 2 Method Number: 1• Analytical sensitivity is defined as the concentration calculated as the mean plus two

standard deviations of replicates of the ARCHITECT Total T3 MasterCheck Level 0 (0.0 ng/mL). The analytical sensitivity (low-linearity) is defined in the ARCHITECT Total T3 assay parameters as 0.25ng/mL.

MasterChecks are no longer manufactured. To verify analytical sensitivity for the ARCHITECT Total T3 assay it is recommended the customer use other suitable samples at the following analyte concentrations:• A sample with no analyte present• A sample with a value that corresponds to a "B" calibrator.Data Reduction

Assay CD-ROM Utilize the current assay CD-ROM which contains ARCHITECT TT3 assay file version 671_007.iae:• List No. 1P60 - i1000SR System Assay CD-ROM - US Special Edition - version 7.0 or

higher• List No. 1P61 - i1000SR System Assay CD-ROM - WW (excluding US) Special Edition -

version 8.0 or higher• List No. 6E58 - i System Assay CD-ROM - US Version 29.0 or higher• List No. 6E59 - i System Assay CD-ROM - WW (excluding US) version 33.0 or higher




Assay Sequence

Keywords Information




Calibration Validity Specifications

The ARCHITECT Total T3 assay parameter file contains 13 checks to insure proper curve adjustments.The ARCHITECT Total T3 calibration uses the following protocol:• When the system scans the reagent kit, it stores the reference curve data.

• The reagent microparticle bottle’s bar code contains the factory-generated reference curve data.

• It contains one RLU value for each of the 6 calibrator points used to generate the reference curve.

• The 2-point calibrator adjustors are run in duplicate.

• The average of these values, in RLUs, are used to adjust the six-point reference curve.• The Total T3 fit curve is set off of the six points using 4PLC, Y-weighted linear

transformation.• The data reduction method is assay specific.• Data reduction math models can be found in Appendix C of the Operations Manual.

• Each six-point calibrator is given a “fit” value, in RLUs. This produces a Fit Curve.• Before this Fit Curve is accepted, it must pass the validity specs as defined in the

parameter file.• These validity specifications ensure the integrity of the cal curve.• The validity specifications used are assay specific.

* All concentrations are ng/mL


Validity Check for… Acceptance value*…

Purpose of the validity check…

Adj 1 RLU % CV < 5.8 % Precision check on Cal 1 to prevent inaccurate curve shape at the low concentration end of the cal curve

Adj 2 Min and Max RLU Deviation from the Average

< 11 % Precision check between the smallest and largest Cal 2 rep to prevent inaccurate curve shape at the highconcentration end of the cal curve

Fit A < 0.25 Checks the adjusted A Cal concentration off of the fit cal curve

Fit B 0.35-0.65 Checks the adjusted B Cal concentration off of the fit cal curve

Fit C 0.7-1.3 Checks the adjusted C Cal concentration off of the fit cal curve

Fit D 1.4-2.6 Checks the adjusted D Cal concentration off of the fit cal curve

Fit E 2.8-5.2 Checks the adjusted E Cal concentration off of the fit cal curve

Fit Adj 1 0.35-0.65 Checks the concentration of the mean Cal 1 off of the fit cal curve




Competitor AssayInfo


ARCHITECT TT3 Bayer ACS:180 T3Bayer ACS: Centaur

Total T3 J&J Vitros Total T3

Method and Format CMIA, 2-step competitive CMIA, competitive CMIA, competitive1-step competitive

Kit Size 100 (1x100), 400 (4x100), 2000 (4x500) 50, 300 80, 400 100

Throughput 200 tests/hour, first result in 29 min 180 tests/hour 240 tests/hour 90 tests/hour

Calibrator Range 0-8 ng/mL 0-8.0 ng/mL 0-8 ng/mL 0-7.8 ng/mL

Calibrators 2 points 2 points 2 points not published

Sample Volume 150 µL 50 µL 50 µL 20 µL

Sample Types serum and plasma serum serum serum/plasma (EDTA, heparin)

Reagent Stability on board: Up to 30 days on board: 8 hours 14 days on board: 8 weeks

Calibration Curve Stability Up to 30 days 7 days 14 days 28 days

Reagent Preparation none none none none

Sensitivity <0.25 ng/mL 0.20 ng/mL 0.1 ng/mL 0.10 ng/mL

Precision

Total CV% At (ng/mL) 3.6-7.0 0.7 2.3-7.3 1.5 2.0-4.2 3.3

Total CV% At (ng/mL) 5.2 0.73 3.4 1.61 3.5 3.20

Total CV% At (ng/mL) 3.44 0.81 2.27 1.78 1.84 3.21

Within Lab CV% At (ng/mL)

8.2 0.47 4.6 1.16 5.3 2.68

Expected Values 0.58-1.59 ng/mL 0.60-1.81 ng/mL 0.60-1.81 ng/mL 0.97-1.69 ng/mL




Competitor Assay Info(continued)

Contents Calibrator:2 Bottles (4 mL each) of ARCHITECT Total T3 Calibrators (Cal 1, Cal 2) prepared in human serum, which has been tested and found to be nonreactive for HBsAg, HIV-1 RNA or HIV-1 Ag, anti-HCV, and anti-HIV-1/HIV-2. Preservative: Sodium Azide.MCC: This product is prepared from human serum with added constituents of human and animal origin, pure chemicals, therapeutic drugs and preservatives. The control is provided in liquid form for convenience.Manual Diluent:1 Bottle (5 mL) of ARCHITECT Total T3 Manual Diluent containing human serum, which has been tested and found to be nonreactive for HBsAg, HIV-1 RNA or HIV-1 Ag, anti-HCV, and anti-HIV-1/HIV-2. Preservative: Sodium Azide.


ARCHITECT TT3 Roche Elecsys T3Bayer Immuno 1 Total

T3 Immulite 2000 Total T3

Method and Format CMIA, 2-step competitive

ECLIA, competition principle

heterogeneous competitive MSA

solid phase, chemiluminescent enzyme immunoassay

Kit Size 100 (1x100), 400 (4x100), 2000 (4x500) 200 125 200, 600

Throughput 200 tests/hour, first result in 29 min

Elecsys 1010: 58 tests/hourElecsys 2010: 88 tests/hour 120 tests/hour 200 tests/hour

Calibrator Range 0-8 ng/mL 0-6.51 ng/mL 0-8 ng/mL 0-6.0 ng/mL

Calibrators 2 points 2 points 6 points adjustors (2 point)

Sample Volume 150 µL 30 µL 15 µL 25 µL

Sample Types serum and plasmaserum/plasma (heparin, EDTA, citrate, sodium fluoride/potassium oxalate)

serum serum

Reagent Stability on board: Up to 30 days on board: 8 weeks on board: 21 days not published

Calibration Curve Stability Up to 30 days

Elecsys 1010: 3 to 7 days (depending on room temperature) Elecsys 2010: One month (same reagent lot), 7 days (same reagent kit on analyzer)

60 days 2 weeks

Reagent Preparation none yes (reconstituted calibrators and controls) must prepare conjugate none

Sensitivity <0.25 ng/mL 0.195 ng/mL 0.06 ng/mL 0.19 ng/mL

Precision

Total CV% At (ng/mL) 3.6-7.0 0.7 2.3-7.3 1.5 2.0-4.2 3.3

Interassay CV% At (ng/mL) 5.4 0.79 4.7 1.81 5.4 3.31

Total CV% At (ng/mL) 13.3 0.46 6.0 1.34 3.9 3.43

Total CV% At (ng/mL) 15 0.73 8.6 0.88 7.0 1.71 5.3 3.62

Expected Values 0.58-1.59 ng/mL 0.8-2.0 ng/mL 0.7-1.4 ng/mL 0.82-1.79 ng/mL




Curve Shape

Curve Stability Calibration curve stability has been verified for up to 30 days.

Evaporation • Due to the impact of humidity, temperature and timing requirements, the minimum volume for a single test is 150 µL.

• To minimize the effects of evaporation, all samples (patient specimens, calibrators, controls, etc.) must be tested within 3 hours of being placed on board the ARCHITECT i System. If the sample is on board for longer than 3 hours, additional sample volume is required.

Frozen Specimen Storage

• Multiple freeze-thaw cycles of specimens should be avoided.• Specimens must be mixed THOROUGHLY after thawing, by LOW speed vortexing or by

gently inverting, and centrifuged prior to use to remove red blood cells or particulate matter to ensure consistency in the results.

Functional Sensitivity

Functional sensitivity is not provided for the ARCHITECT Total T3 assay. Refer to analytical sensitivity.

Intended Use The ARCHITECT Total T3 assay is a Chemiluminescent Microparticle Immunoassay (CMIA) for the quantitative determination of total triiodothyronine (Total T3) in human serum and plasma.


ARCHITECT Total T3 Curve Shape

0

5 0000

10 0000

15 0000

20 0000

25 0000

0 2 4 6 8 1 0

Calibra tor Co ncentra tion (ng/mL )

Rel

ati

ve L

ight

Uni

ts (

RL

Us)




Least Squares Correlation Plot(s)

Correlation of ARCHITECT Total T3 vs. AxSYM Total T3Accuracy by Correlation

Least Squares Linear Regression Method

Levey-Jennings Utilize the following to set-up the Manufacturer’s Expected Mean and S.D. for the Abbott Immunoassay MCC’s:

• Refer to the MCC Package Insert and lot specific data sheet as a foundation to establish your own expected mean and 1 S.D.

• Run the MCC until you have sufficient data to set your lab’s control ranges. Refer to the keyword “Setting Control Ranges” as a guideline.





Matrix Effects • No matrix effects are known or have been observed for this assay.• Matrix Definition: Interference caused by undefined actors in biological fluids. Different

samples each containing the same amount of analyte may give different results. The lower the proportion of sample, the overall assay reaction, the less influence these matrix effects have on accuracy of the assay.

(Wild, David. The Immunoassay Handbook. Stockton Press, 4)Microbial Contamination

The effect of microbial contamination of samples has not been determined.

Onboard Stability

• The ARCHITECT Total T3 assay reagent kit may be stored onboard the ARCHITECT i System for a maximum of 30 days. After 30 days the reagent kit must be discarded.

• For information on tracking onboard time, refer to the ARCHITECT i System Operations Manual, Section 5.

Panel Requirements

The following table represents the most restrictive sample requirements for the Thyroid panel assays, which include, but are not limited to:


Specimen Type(s): • Human serum • Plasma

Collection Tube(s) • Serum Separator• Sodium Heparin• Lithium Heparin• Potassium EDTA anticoagulant

Storage/Sample Handling

• 2-8°C for 6 days• Can freeze samples at -10°C for up to 6 days• Avoid multiple freeze-thaw cycles of

specimensPotential Interferences

• Hemoglobin: <10% at 500 mg/dL• Bilirubin: <10% at 20 mg/dL• Triglycerides: <10% at 2000 mg/dL• Protein: <10% at 2 and 12 g/dL




Passing-Bablok Correlation Plot(s)

ARCHITECT Total T3 versus AxSYM Total T3Accuracy by Correlation

Passing-Bablok Linear Regression Method

Reagent Appearance

Antifoam, which may look like a floating layer on top of the reagents (similar in appearance to oil on top of water), does not affect assay performance or results. Reagents may be run without additional mixing.





Reference Assay Sequence

The AxSYM Total T3 reagents and sample are pipetted in the following sequence:SAMPLING CENTER• Sample and all AxSYM Total T3 reagents required for one test are pipetted by the

Sampling Probe into various wells of a reaction vessel (RV).The RV is immediately transferred into the Processing Center. Further pipetting is done in the Processing Center by the Processing Probe.PROCESSING CENTER• Sample and Anti-T3 Coated Microparticles are combined in one RV well.• The free (unbound) T3 in the sample binds to the Anti-T3 Coated Microparticles forming

an antibody-antigen complex.• An aliquot of the reaction mixture, containing the antibody-antigen complex bound to the

microparticles, is transferred to the matrix cell. The microparticles bind irreversibly to the glass fiber matrix.

• The T3: Alkaline Phosphatase Conjugate is dispensed onto the matrix cell and binds to the available sites on the Anti-T3 Coated Microparticles.

• The matrix cell is washed to remove unbound materials.The substrate, 4-Methylumbelliferyl Phosphate, is added to the matrix cell and the fluorescent product is measured by the MEIA optical assembly.

Reference Method

Abbott AxSYM System Microparticle Enzyme Immunoassay (MEIA) (List 7A52).

Reportable Range Verification

• Group Number: 2 Method Number: 2• Refer to the ARCHITECT i System Operations Manual, Appendix B.MasterChecks are no longer manufactured. To verify reportable range CLIA Regulation 493.1217 requires a minimum of three points using suitable samples that span the assay range as follows:• A sample with no analyte present• A sample with a value set near the mid point or clinically relevant area of the assay

dynamic range• A sample with a value set at or below the upper calibration range (value set to ensure the

customer can claim the full reportable range of the assay).Use suitable samples at the concentrations below:

“Same As” List NOTE: This information is being supplied for troubleshooting and trend analysis.The following list numbers represent the same formulation as the ARCHITECT Total T3 assay Calibrators and Controls; however, they are not intended for use on theARCHITECT i System, and may NOT be used in the ARCHITECT Total T3 assay.The ARCHITECT Total T3 Calibrators (7K64-01) represent the same formulation as:• AxSYM Total T3 Calibrators (List 7A52-02)• IMx Total T3 Calibrators (List 2250-02)


Sample Type ConcentrationNo analyte 0 ng/mL

Clinically Relevant Area ~ 1.5 ng/mL

Upper calibration range ~ 8.0 ng/mL (Cal 2)




Setting Control Ranges

Follow the NCCLS guideline for precision performance evaluation. (National Committee for Clinical Laboratory Standards, Evaluation of Precision Performance of Clinical Chemistry Devices - Approved Guideline. NCCLS Document EP5-A. Wayne, PA: NCCLS, 1999.)• Use 20 replicates for each level.• Generate each replicate on a separate assay run conducted across all modules.• Complete analysis for every analyte.• Calculate the mean and standard deviation (SD) of the replicates.• Use the SD to set the SD range around the target value.• Recommend 2 to 3 SD range.• Determine value by laboratory’s quality control policies.

Shipping Conditions

* Multi-constituent Controls


Sample • When shipped, specimens must be packaged and labeled in compliance with applicable state, federal and international regulations covering the transport of clinical specimens and infectious substances. Prior to shipment, it is recommended that specimens be removed from the clot, serum separator or red blood cells.

Reagent Kits • Ambient

Control Kits • N/A

Calibrator Kits • Ambient

Immunoassay - MCC*(liquid, 6E20-10)

• Frozen on dry ice




Specimen Handling

• Human serum (including serum collected in serum separator tubes) or plasma collected in sodium heparin, lithium heparin, or potassium EDTA anticoagulant tubes may be used in the ARCHICTECT Total T3 assay. Other anticoagulants have not been validated for use with the ARCHITECT Total T3 assay. Follow the manufacturer’s processing instructions for serum or plasma collection tubes.

• When serial specimens are being evaluated, the same type of specimen should be used throughout the study.

• The ARCHITECT i System does not provide the capability to verify specimen type. It is the responsibility of the operator to verify the correct specimen types are used in the ARCHITECT Total T3 assay.

• Use caution when handling patient specimens to prevent cross contamination. Use of disposable pipette tips is recommended.

• Do not use heat-inactivated specimens.• For optimal results, inspect all samples for bubbles. Remove bubbles with an applicator

stick prior to analysis. Use a new applicator stick for each sample to prevent cross contamination.

• For optimal results, serum and plasma specimens should be free of fibrin, red blood cells or other particulate matter.

• Ensure that complete clot formation in serum specimens has taken place prior to centrifugation. Some specimens, especially those from patients receiving anticoagulant or thrombolytic therapy may exhibit increased clotting time. If the specimen is centrifuged before a complete clot forms, the presence of fibrin may cause erroneous results.

Standardization The calibrators are manufactured using L-Triiodothyronine, sodium salt, (USP Grade) and signal matched to Abbott internal reference standards, which are traceable to the USP Reference Standard L-Triiodothyronine (free acid) at each concentration level. The concentration of the T3 stock solution used to make the internal reference standards is determined by HPLC.

Storage

* If not indicated otherwise in the table, all listed components can be used until they reach their expiration date (if stored and handled as directed).

Special Handling Instructions:1. Reagents may be stored on board the ARCHITECT i System for up to 30 days. If

removed, they need to be stored in an upright position with septum and replacement cap installed.NOTE: If the microparticle bottle does not remain upright (with a septum installed) while in refrigerated storage off the system, the reagent kit must be discarded.

2. Gently swirl all calibrators and controls before use to ensure homogeneity of material.


Type Condition Storage * Use

Reagents1 Unopened/Opened 2-8°C Immediate

Controls2 N/A N/A N/A

Calibrators2 Unopened/Opened 2-8°C Immediate

Immunoassay - MCC(6E20-10, Liquid)

UnopenedUnopened

Opened

-20°C or colder2-8°C for 7 days2-8°C for 7 days

Allow to reach 2-8°C




Summary and Explanation of Test

T3 levels are also affected by conditions which affect TBG concentration.13-15 Slightly elevated T3 levels may occur in pregnancy or during estrogen therapy, while depressed levels may occur during severe illness, malnutrition, in renal failure and during therapy with anti-thyroid drugs, propranolol and propylthiouracil and salicylates.2,16,17 In patients with severe or chronic illnesses, many abnormalities of thyroid hormone balance occur. T4 production and the extent of serum thyroid hormone binding may be independently abnormal, resulting in a low, normal or high free T4 estimate. Serum T3 concentrations are often low; TSH levels may be normal or slightly elevated. Total T3 measurements may be valuable when hyperthyroidism is suspected and the free T4 estimate is normal.13

The ARCHITECT Total T3 assay is to be used as an aid in the assessment of thyroid status.

Westgard Rules Instrument software contains ARCHITECT specific Westgard rules. All rules apply to the Total T3 assay. Westgard rules are configured by the customer. Refer to the ARCHITECT i System Operations Manual, Section 2, for further information.





Package Insert Bibliography1. Budavari S, editor. Merck Index (11th Ed.). Rahway, NJ: Merck and Co., Inc., 1989:868.2. Larsen PR. Triiodothyronine: Review of Recent Studies of Its Physiology and Pathophysiology in Man.

Metabolism 1972;21:1073-1092.

3. Ekins RP, editor. Methods for the Measurement of Free Thyroid Hormones. Amsterdam: Excerpta Medica Foundation. 1979;72-92.

4. Robbins J, Rall JE. The Iodine-containing Hormones. In: Hormones in Blood (3rd Ed.). London: Academic Press, 1979:1:632-667.

5. DeGroot LJ, Larsen PR, Refetoff S, Stanbury JB. Transport of Thyroid Hormone and Cell Uptake. In: The Thyroid and Its Diseases. New York: Wiley and Sons, 1984;62-66.

6. Wahner HW, Gorman CA. Interpretation of Serum Tri-Iodothyronine Levels Measured by the Sterling Technic. N Engl J Med 1971;2824:225-230.

7. Marsden P, McKerron CG. Serum Triiodothyronine Concentration in the Diagnosis of Hyperthyrodism. Clin Endocrinol 1975;4:183-189.

8. Ivy HK, Washner HW, Gorman CA. Triiodothyronine (T3) Toxicosis: Its Role in Graves’ Disease. Arch Intern Med 1971;128:529-534.

9. Hollander CS, Mitsuma T, Nihei N, Shenkman L, Burday SZ, Blum M. Clinical and Laboratory Observations in Cases of Triiodothyronine Toxicosis Confirmed by Radioimmunoassay. Lancet 1972;1:609-611.

10. Sterling K, Refetoff S, Selenkow HA. T3 Thyrotoxicosis: Thyrotoxicosis Due to Elevated Serum Triiodothyronine Levels. JAMA 1970;213:571-575.

11. Hollander CS, Mitsuma T, Shenkman L. Stevenson C, Pineda G, Silva E. T3 Toxicosis in an Iodine-Deficient Area. Lancet 1972;2:1276-1278.

12. Ermans AM. Disorders of Iodine Deficiency. In: Ingbar SH, Braverman LE, editors. The Thyroid (5th Ed.). Philadelphia: JB Lippincott Co., 1986:705-721.

13. Kaplan MM, Larsen PR, Crantz FR, Deau VJ, Rossing TH. Prevalence of Abnormal Thyroid Function Test Results in Patients with Acute Medical Illnesses. Am J Med 1982;72:9-16.

14. Bermudez F, Surks MI, Oppenheimer JH. High Incidence of Decreased Serum Triiodothyronine Concentration in Patients with Nonthyroid Disease. J Clin Endocrinol Metab 1975;41:27-40.

15. Oppenheimer JH. Thyroid Function Tests in Nonthyroidal Disease. J Chronic Dis 1982;35:697-701.

16. Abuid J, Larsen PR. Triiodothyronine and Thyroxine in Hyperthyrodism: Comparison of the Acute Changes During Therapy with Antithyroid Agents. J Clin Invest 1974;54:201-208.

17. Felig P, Baxter JD, Broadus AE, Frohman LA, editors. Endocrinology and Metabolism (2nd Ed.). New York: McGraw-Hill Book Co., 1987:408-416.

18. US Department of Labor, Occupational Safety and Health Administration, 29 CFR Part 1910.1030, Bloodborne pathogens.

19. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories. 5th ed. Washington, DC: US Government Printing Office; January 2007.

20. World Health Organization. Laboratory Biosafety Manual. 3rd ed. Geneva:World Health Organization; 2004.

21. Clinical and Laboratory Standards Institute. Protection of Laboratory Workers from Occupationally Acquired infections: Approved Guideline - Third Edition. CLSI Document M29-A3. Wayne, PA: Clinical and Laboratory Standards Institute; 2005"

22. National Committee for Clinical Laboratory Standards, Evaluation of Precision Performance of Clinical Chemistry Devices – Approved Guideline. NCCLS Document EP5-A. Wayne, PA: NCCLS, 1999.




23. Passing H, Bablok W. A New Biometrical Procedure for Testing the Equality of Measurements from Two Different Analytical Methods. J Clin Chem. Clin Biochem. 1983;21:709-20.

ARCHITECT, AxSYM, IMx and MasterCheck are trademarks of Abbott Laboratories in various jurisdictions.Triton is property of its respective owner.




Revision History

Part Number RevisionDate

(Month, Year)Pages Revised

and Added Prepared by:

201422-101 1 June, 2005 Complete Rewrite Creative Network

201422-102 2 July 2010Cover page, 1, 2, 6, 9-16, 20-22, 26

of 26GSS ePublishing Dallas


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