Presents

Sensi-Ring™ Antibiotic Sensitivity Detection System

Procedures and Techniques For Performing

In-house Antibiotic Sensitivities On Cultured

Kacey MultiChrome™ Bi-pates.

“Choosing the right drug for the right bug”

Why use Kacey Sensi-Ring™?

In house rapid identification (24 hours) of bacterial antibiotic

sensitivity using 8 of the most common veterinary antibiotic

choices per ring.

Cost Effective: Less than 1/3 the cost of a commercial laboratory

like Antech and Idexx.

Specific Sensi-Ring for both Gram Positive and Gram Negative

System specific Sensi-Ring:

- Urinary Tract Infection

- Ear Infection

- Skin and Wound Infection

Sensi-Ring™ Supplies For Each Test

For added convenience we also offer a Culture & Sensitivity Starter Kit

enough for four complete tests with all the supplies you need. Product #20241

Step 1: Obtaining live cultured bacteria

from a Kacey MultiChrome™ bi-plate.

• Using the yellow loop, take 3 or 4 passes through

the bacterial growth on the MultiChrome™.

For bacterial colonies in both chambers see next slide.

Step 1 (con’t): Procedure For MultiChrome

bi-plates with bacteria on both the Gram

Positive and Negative side.

• In this case, you will prepare two Kacey Maxi Mueller

Hinton plates as indicated. One for the (+) and one for

the (-).

• To each MH plate you will add the appropriate Sensi-Ring

disk Example: place a Gram (+) in the MH plate with Gram (+) and

Gram (-) in the MH plate with Gram (-). Then incubate each.

Step 2: Mixing with Saline (WST) to dilute

• Insert the yellow loop into the saline and twirl

to make a cloudy solution. This is an important

step to avoid overgrowth of a bacterial colony.

Step 3: Removing the diluted sample with

enclosed Rayon* Swab

• Take a sample using the applicator tip of the swab.

* Do not use cotton swabs even if sterile. Cotton will release

a chemical that will alter the results.

Step 4: Inoculate diluted bacteria onto

Kacey Maxi* Mueller Hinton plate

• Using the diluted bacteria solution on the soaked

swab, evenly distribute the sample onto a warmed

Kacey Maxi Muller Hinton plate (37 degrees 15-20 min).

• Streak the plate as shown

below. 11-5, 1-7, 9-3 o’clock

*Kacey Maxi Mueller Hinton plates are “true” 100mm plates. Most

commercially sold Muller Hinton plates are 88mm and will not work

with this system.

Step 5: Apply the Sensi-Ring™

• Pick the appropriate Sensi-Ring™ ie. Gram Positive,

Gram Negative, UTI or Skin/wound depending on

samples origin.

• Grasp the tab on the inside of the ring and lower

onto the agar upside down with lettering facing agar,

centering the ring with tweezers.

Step 6: Tamp down Sensi-Ring™

• Make sure that all of the disks touch the agar

plate to allow antibiotic diffusion.

Step 7: Incubate Upside Down

• Place Kacey Maxi Mueller Hinton plate now

inoculated upside down to prevent any contamination.

• Place into a preheated Kacey incubator at 37 degrees

for 24 hours.

24 hours

Step 8: Remove and read after 24 hours

• Carefully remove plate from incubator keeping it

upside down so that Sensi-Ring letters can be seen.

• Use gloves. Remember this is a live bacterial colony.

Step 9: Use the Kacey Zone Reader Card

• Place Kacey Maxi Mueller Hinton Plate upside down

with antibiotic letters facing up.

• Place Zone Reader over the plate and align the

notch of the Reader to the notch of the Sensi-Ring.

• Concentric circles should now encircle all of the disks.

• Read the Inhibition Zone in mm for each disk.

Notch

Step 10: Log Test Results

• Using the Patient Lab Report form supplied log

inhibition zones for each antibiotic disk.

Step 11: Treat with appropriate antibiotic

• Based on results, a course of action can now be

taken with the most efficacious antibiotic(s).

• The following is a list of commonly found Aerobic

Bacteria in small animal practice:

Gram Positive: Staphylococci, Streptococci, Enterococci

Gram Negative: E coli, Proteus, Klebsiella, Pseudomonas

Ear: Staphylococci, Pseudomonas, Proteus, E. coli, Enterococci, Streptococci

UTI: Staphylococci, Proteus, E. coli, Klebsiella, Enterococci, occasionally Pseudomonas

Skin & Wound: Staphylococci, Streptococci, Pseudomonas, Enterococci, Proteus

Step 12: Disposal of plate & clean up

• Once read, plate should be sprayed with a bactericidal

agent, covered with its lid taped shut, placed in a plastic

bag and disposed of in your bio-hazard bin.

• If you have a Kacey Sanitizer unit follow its instructions.

Place lid back on the plate, tape it shut, bag it and dispose

of it in your bio-hazard bin.

• Dispose of all used supplies in your bio-hazard bin.

• Spray your counters down and wash your hands in an

aseptic manor. Remember, these are live cultures.

Alternate Bacteriology Protocol Inoculating

Mueller Hinton Plate First Without Using the

MultiChrome Culture Plates

With Kacey Culture and Sensitivity system is it possible to

first directly inoculate a Maxi Mueller Hinton plate, incubate

for 24 hours and determine most efficacious antibiotic before

actually identifying the pathogenic bacteria.

A sample can then be drawn from the center area of the

Sensi-Ring™ on the Maxi Mueller Hinton plate and

transferred to a MultiChrome™ plate for incubation and

identification of the bacteria(s).

• Urine Samples: Spin down urine and do cytology on

the sample of the sediment.

- If bacteria is present, directly inoculate sediment

onto a Kacey Maxi Mueller Hinton plate with a

sterile Rayon swab enclosed smeared evenly

over the surface as described in step 4 .

• Ear Samples: Do a cytology on each ear swab.

- If bacteria is present, directly inoculate the Kacey

Maxi Mueller Hinton plate with sample smeared

evenly over the surface as described in step 4.

Alternate Bacteriology Protocol Inoculating

Mueller Hinton Plate First Without Using the

MultiChrome Culture Plates

Sensi-Rings™ case samples

Case study: 2 year old canine which cultured on the MultiChrome™ both a

large colony of Gram Positive Staph in pink and a smaller Strep colony in blue.

The colonies when tested for sensitivity using the Sensi-Ring™ indicated

sensitivity to a variety of antibiotics not previously used.

Staph

Strep

UTI and Ear samples

Sensi-Rings™ Ability to identify

different strains of bacteria.

• Sensi-Rings™ used to identify antibiotic sensitivity

on different strains of cultured Staphylococci.

Sensi-Rings™ Quick Reference Card

Kacey Diagnostics Quick Reference For Culture & Sensitivity Study

INNOCULATION OF CULTURE BI-PLATE

1) Remove the plate from refrigerator & pre-heat

in the incubator @37 degrees for 15-20 minutes

prior to inoculating the Kacey MultiChrome bi-

plate.

2) Using a sterile inoculating loop ( 20uL) for

liquids and a Kacey Sterile Swab for solids

,inoculate the sample onto both sides of the

Kacey Multichrome

Bi-plate, by utilizing a zigzag streaking motion.

3) Replace clear lid and place Kacey MultiChrome

Bi-plate in the incubator upside down (inverted

position) and incubate at 37°C +/- 2 degree C for

no less then 24 hours . The plate should be

examined after 24 hours, but no later than 48

hours after incubation.

INOCULATION OF MULLER HINTON (MH) PLATE

1) Using the Kacey Swab containing the now diluted

WST dilution streak the 1st time the entire periphery

of the Mueller Hinton plate making a 360° circle

2) Streak a 2nd time again the Muller Hinton Plate

with a fresh sample of the WST dilution usingbroad

strokes starting at theto 3 o’clockposition.

2) Streak a 3rd time again the Muller Hinton plate

with a fresh sample of the WST dilution using wide

broad strokes starting at the 11 to 5 o’clock position

3) Remove “Sensi-Ring” from the foil pouch with

tweezers at the inner tab & place the “Sensi-Ring”

on to MH plate tapping down in non-disk areas, label

the specimen to be incubated.

4) Place the MH Inoculated plate into an incubator

upside down, set timer & incubate for 24 hours.

Remove & read inhibition zones with Kacey Digital

Reader.

1 2 3 TRANSFER TO MULLER HINTON PLATE USING

KACEY “WST” TUBES

1) Taking a Kacey Sterile Swab carefully dab on to

three (3) different places containing the bacteria on

the MultiChrome Bi-plate.

2) Immediately place sterile Kacey Swab containing

the bacteria into a Kacey Working Solution tube

(WST containing 1.0 ml of 0.085%solution).

3) Mix the Kacey Swab with a gentle twirling motion

while in the WST tube for approximately 3-5

seconds. Use the same Kacey Swab for

inoculation of the Muller Hinton plate in step #3.

Quick reference only. Refer to detailed instruction manual for additional information.

GN

GP

1

2

3

WST Dilution

Rotate Swab 3-5 sec

GN

GP

24 Hrs 24 Hrs

New Kacey Sterile Swab

Kacey Diagnostics

MICROBIAL ZONE INHIBTION

INTERPRETIVE CHART* w/ Zone Diameter Interpretive Standards for

common veterinary Antibiotics

Amikacin …………………………………………….

Amoxicillin …………………………………………..

Amoxi Clavulante …………………………………..

Augmentin-(Clavamox)…………………………….

Azithromycin-(Zithromax)………………………….

Cephalexin-(Keflex)………………………………..

Cefapodoxime ……………………………………..

Ceftazdime ………………………………………….

Ciprofloxacin –(Cipro)……………………………..

Clindamycin –(Clincaps)…………………………..

Cotrimoxazole (trimethoprim/sulfa)………………

Choloramphenicol-(Chloromycetin)………………

Cefalothin-(Keflin)………………………………….

Doxycycline-(Vibramycin)…………………………

Enrofloxacin-(Baytril)……………………………….

Gentamycin-(Gentocin)…………………………….

Neomycin …………………………………………...

Oxacillin ……………………………………………..

Penicillin……………………………………………..

Polymycin …………………………………………..

Ticarcillin-(Ticar)…………………………………….

Tobramycin-(Tobrex)……………………………….

Trimethoprim/Sulfa-(Bactrim Septra Primor)…….

30ug

30ug

30ug

30ug

13ug

30ug

10ug

30ug

5ug

30ug

25ug

30ug

30ug

30ug

5ug

10ug

30ug

1ug

10ug

300ug

75ug

10ug

25ug

=14

=13

=13

=13

=13

=14

=17

=14

=15

=13

=10

=17

=14

=12

=17

=6

=12

=10

=28

=8

=14

=12

=10

15-16

14-17

14-17

14-17

14-17

15-17

18-20

15-17

16-20

14-17

11-15

18-20

15-17

13-15

18-20

7-9

13-16

11-12

——

9-11

15-19

13-14

11-15

=17

=18

=18

= 18

= 18

= 18

=21

=18

= 18

= 19

= 16

= 21

= 18

= 16

= 21

= 10

=17

=13

= 29

=12

= 20

= 15

= 16

AK

A

AMC

AUG

ATH

CFX

CPD

CAZ

CIP

CD

TS

C

KF

DXT

ENF

GM

NE

OX

PG

PB

TC

TN

T/S

Antimicrobial Agent Name Sensi Code Disc Potency Resistant Intermediate Susceptible

*These are the most widely accepted standard

procedures used by the US Food and Drug

administration (FDA) and the World Health

Organization (WHO). These standard procedure

values were adopted as a consensus standard by the

Clinical and Laboratory Standard Institute (CLSI)

formerly known as NCCL and are periodically

updated. The latest CLSI documentation should

always be consulted for current recommendations.

The list of abbreviations for the antibiotics has been

selected as standardization by KACEY® for

worldwide sales, distribution, nomenclature and

identification of the Kacey Sensi-Rings.

It is highly recommended that any bacterial agent

whose etiology and treatment is beyond the scope

of this system be sent to a qualified veterinary

reference lab for more detailed testing. It is also

recommended that any sample sent the lab be

obtained from the middle of the Sensi Ring on the

Kacey Maxi Muller Hinton Plate".

Sensi-Rings™ & MultiChrome™ are trademarks Kacey® © 2013 Rev 10.3.2013 All Rights Reserved

Sensi-Rings™ Quick Reference Card (con’t)

1854 A Hendersonville Rd

Asheville, NC 28803

Ph: 1(828)685-3569

Fax: 1(828)685-3571

www.kaceyinc.org

“Kacey Products Providing Better Animal Care”

Kacey Diagnostics © 2014 All Rights Reserved

Rev 12.30.12

“MultiChrome”, “Sensi-Rings” are trademarks of Kacey® Inc.