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ImmunoComb Feline VacciCheck

This page contains information on ImmunoComb Feline VacciCheck for veterinary use.
The information provided typically includes the following:
  • ImmunoComb Feline VacciCheck Indications
  • Warnings and cautions for ImmunoComb Feline VacciCheck
  • Direction and dosage information for ImmunoComb Feline VacciCheck

ImmunoComb Feline VacciCheck

This treatment applies to the following species:
Company: Aventix

FELINE PANLEUKOPENIA VIRUS ANTIBODY TEST KIT, DOT BLOT

INSTRUCTION MANUAL

December 31, 2015

In Vitro Use Only

Produced by: Biogal Galed Labs. - Galed, 1924000 - Israel

Tel. 972-4-9898605, Fax. 972-4-9898690

Produced for: Biogal USA, Inc. - 28300 DuPont Hwy., Millsboro, DE 19966 - USA

Tel. (302)934-8700, Fax. (302)934-8745

Distributed by: Aventix Animal Health - Burlington, ON Canada L7L 5R7

Tel. 1-877-909-2242, Fax. (905) 332-4844

U.S. Permit No. 454A

I. INTENDED USE OF THE KIT

This kit is used for the detection of antibodies to Feline Panleukopenia virus (FPLV) in cat whole blood or serum.

II. GENERAL INFORMATION

Feline Panleukopenia Virus (FPLV), is recognized as an important cause of illness and death in cats. Kittens are more susceptible to FPLV especially after weaning when protective Maternally Derived Antibody (MDA) levels decrease.

Sometimes MDA may actually interfere with vaccinations that are given for immunization.

In many countries, vaccination programs have significantly curtailed, but not eliminated the incidence of this disease. Thus, FPLV continue to be of a great clinical concern among veterinarians worldwide and still present a diagnostic challenge.

III. DESCRIPTION OF DISEASES

Feline Panleukopenia (FPLV also known as Feline Distemper or Feline infectious enteritis) is a highly contagious viral disease that can kill both kittens and unvaccinated adult cats. Symptoms include sudden onset of fever, lack of appetite, dehydration, depression, vomiting and dizziness. Infected cats may show a decreased number of whole blood cells.

Veterinarians typically make a presumptive diagnosis of Feline Panleukopenia based on clinical signs, which range in severity from mild to severe. Laboratory tests can be helpful for confirming the diagnosis. In addition to hematology and blood chemistry, serology is becoming a more widely accepted diagnostic tool.

IV. WHAT IS THE IMMUNOCOMB ASSAY?

The ImmunoComb is a modified ELISA, which can be described as an enzyme labeled “dot blot assay” that detects antibodies in serum or whole blood.

The kit contains all the necessary reagents for developing the test. Results for the Feline Panleukopenia test are obtained within 21 minutes.

V. HOW DOES THE IMMUNOCOMB WORK?

● The ImmunoComb Kit contains 2 main components: a comb shaped plastic card, hereafter referred to as the Comb and a multi compartment developing plate.

● The Comb has 12 teeth - sufficient for 12 tests. Each tooth will be developed in a corresponding column of wells in the developing plate. Individual or multiple tests are processed by breaking off the desired number of teeth from the Comb.

● Test spot of FPLV is attached to each tooth on the Comb. The upper spot is a Positive Reference.

● Purified FPLV antigen is attached to the lowest spot. (See figure in section X).

● Specific antibodies, if present in the sample used in row A of the multicompartment developing plate, react with the FPLV antigen on the Comb test spot.

● At the end of the developing process, described in section VII, positive result will be indicated by purple-grey colored spot developed via enzymatic reaction.

● On each tooth of the developed Comb you should see the upper Positive Reference spot. Test spots of FPLV may appear, depending on the result.

● Results are scored using the Positive Reference spot and CombScale (see section IX).

VI. KIT CONTENTS

The 12-tests kit

The ImmunoComb Card - One Comb sufficient for 12 tests.

Developing Plate - One plate, divided into compartments A-F, that are subdivided into 12 wells. The plate compartments are pre-filled with the reagent solutions.

Disposable Tweezers - For piercing the foil cover of the developing plate compartments.

CombScale - One calibrated CombScale color card for scoring reaction intensities.

Heparinized Capillary Tubes - A vial with twelve capillary tubes and a wire piston, intended for transferring whole blood or serum specimen into compartment A of the developing plate. The tubes are calibrated for dispensing 5 microliters serum (lower band) or 10 microliters blood (upper band).

Documentation - Instruction manual with detailed instructions.

The 120-tests kit

The ImmunoComb Card - Ten Combs sufficient for 120 tests.

Developing Plate - Ten plates, each divided into compartments A-F, that are subdivided into 12 wells. The plate compartments are pre-filled with the reagent solutions.

Disposable Tweezers - For piercing the foil cover of the developing plate compartments.

CombScale - One calibrated CombScale color card for scoring reaction intensities.

Documentation - Instruction manual with detailed instructions.

VII. STEP BY STEP WITH THE IMMUNOCOMB

Before conducting the test, bring the developing plate to room temperature by removing all kit components from the kit carton and place them on the work bench for 60-120 minutes or incubate only the plate at 37°C/98.6°F for 25 minutes.

Perform assay at room temperature of 20° - 25° C / 68° - 77° F).

(1) Obtain blood sample from cat.

(2) Mix reagents by gently shaking the developing plate several times prior to use.

Use the tweezers to pierce the protective aluminum cover of row A. One well for each sample/specimen.

(3) Use a pipette or a capillary tube.

For testing whole blood use 10µl.

For testing serum use 5µl.

(4) Deposit a sample into a well in row A. Raise and lower pipette/piston plunger several times to achieve mixing.

Avoid spillage and cross-contamination of solutions.

Do not open any well of row A or other rows which you do not intend to use.

Do not remove aluminum cover of developing plate all at once.

Remove the Comb from its protective wrapping. Do not touch the teeth of ImmunoComb card. For testing less than 12 samples, cut or break the Comb by folding in allocated notches for the number of tests required.

Note: Mixing during incubation according to instructions is critical for valid results.

**To improve mixing, move the Comb up and down 3-4 times. During incubation, repeat the same mixing process 2-3 times.

Avoid scratching the front active side of the Comb by leaning it to the back while mixing.

Gently shake off excess liquid from Comb teeth onto a tissue before moving it to the next row.

● Insert the Comb into the open well(s) in row A (printed side facing you) and incubate for 5 minutes. Mix as described above.**

● Use tweezers to pierce the foil of the next well(s) in row B. Shake off excess liquid and insert Comb for 2 minutes. Mix as described above.**

● Pierce the foil of the next well(s) in row C. Shake off excess liquid and insert Comb for 5 minutes. Mix as described above.**

● Pierce the foil of the next well(s) in row D. Shake off excess liquid and insert the Comb for 2 minutes. Mix as described above.**

● Pierce the foil of the next well(s) in row E. Shake off excess liquid and insert the Comb for 2 minutes. Mix as described above.**

● Pierce the foil of the next well(s) in row F. Shake off excess liquid and insert the Comb for  5 minutes. Mix as described above.**

● Upon completion of the color development in row F, move the Comb back to row E for 2 minutes for color fixation. Take the Comb out and let it dry for 5 minutes before reading the results.

VIII. READING AND INTERPRETING THE IgG ANTIBODY RESULTS

● The upper spot should give a distinct purple-grey color. This is the same color tone that is generated by a positive response of anti FPLV antibodies at 1:80 H.I. When using the CombScale, this spot should be read as S3 (see section IX).

● The lower spot on the Comb gives the result of FPLV IgG antibodies in the specimen.

● Compare the color tone of FPLV test spot with the Positive Reference spot. (See section IX).

● A color tone equal or darker (S3-S6) than the reference spot is considered a positive response (HI titer ≥ 1:80 for FPLV).

● A faint color tone of S1 or S0 is considered a negative result.

● A color tone that matches with S2 is considered a weak positive result.

● A test spot with a washed blue appearance is invalid. Refer to Biogal for further advice.

IX. READING RESULTS WITH THE COMBSCALE

The CombScale S value is the number that appears in the yellow window corresponding to the color tone, when Positive Reference color is calibrated to S3.

When the Comb is completely dry, align it with the calibrated color CombScale provided in the kit. Find the tone of purple-grey on the CombScale that most closely matches the Positive Reference spot (upper spot). Slide the yellow ruler until the C+ mark appears in the window above that color you just found.

Hold the ruler in this position during the entire reading. This step actually calibrates the C+ to S3, which is the “cut-off” point to which test spots will be compared.

Find the tone of purple-grey on the CombScale that most closely matches the test result spot (lower spot). The number that appears in the window above is the Comb score (S0-S6).

Repeat this step with each test spot, separately.

Another way to read the results is by using the CombScan, version 2.2.0. This is a software program that utilizes a computer and a TWAIN compatible scanner. When a Comb is placed on the scanner, the program translates the color results into numerical values. The CombScan assists labs in reading ImmunoComb results and conserving the data, and is supplied free of charge upon request. See CombScan insert for complete instructions on validating and operating the program.

X. EXAMPLE OF A DEVELOPED COMB

Tooth No.

Results of Panleukopenia

Remarks

1

S0

Negative

 

2

S4

Positive

 

3

≥S5

High Positive

 

4

S0

Negative

 

5

≥S3

Positive

 

6

S0

Negative

 

7

S2

Weak Positive

 

8

 

Invalid

No Positive Reference. Repeat test.

9

 

Invalid

High Background. Repeat test.

10

S0-S1

Negative

 

11

≥S3

Positive

 

12

≥S3

Positive

High Background with positive result.

XI. LIMITATIONS OF THE KIT

● The Panleukopenia Virus ImmunoComb antibody test kit is subject to limitations inherent in any serologic assay, including false negative or false positive results. A false negative result occurs when the specific antibody is below the level detected by the test. This is most likely to happen when cats are tested shortly after the onset of infection/vaccination.

● As is the case for other serologic methods, Panleukopenia serum IgG antibodies are not dectectable by the ImmunoComb assay until one to two weeks post virus exposure. A false positive result can occur in the ImmunoComb assay if substantial amount of antibodies in a specimen bind non-specifically to an antigen spot for reasons that are not clearly understood.

● False positive as well as non-specific (NS) results may occur because of poor quality due to improper sample collection, preparation and/or storage. It is strongly recommended that NS and questionable results be corroborated by retesting specimens with the ImmunoComb test and alternate diagnostic methods.

XII. STORAGE & HANDLING

1. Store the kit under normal refrigeration (2° - 8° C / 36° - 46° F). Do not freeze the kit.

2. Do not mix reagents from different kits or from different compartments of the same kit.

3. The ImmunoComb kit contains inactivated biological material. The kit must be handled and disposed of in accordance with accepted sanitary requirements.

XIII. REFERENCES

AAHA Vaccine Task Force. (2006). JAAHA, 42, 80-89.

DiGangi et al. (2011). J Feline Med Surg. 13(12): 912-918.

Lappin et al. (2002). J Am Vet Med Assoc. 220(1), 38-42 .

Mouzin et al. (2004). J Am Vet Med Assoc. Jan 1, 224(1), 61-66.

Waner et al. (2006). J Vet Diag. Invest. 18. (3), 267-270.

For further assistance please contact:

Biogal Galed Labs Acs. Ltd., Galed, 1924000, Israel Tel: 972-4-9898605 - Fax: 972-4-9898690.

E-mail: info@biogal.co.il

Website: www.biogal.co.il

Instruction Cat. No: 63CVV711

CPN: 1378025.0

AVENTIX ANIMAL HEALTH CORP.
4350 MAINWAY, BURLINGTON, ON, L7L 5R7
Telephone:   905-332-4744
Toll-Free:   877-909-2242
Fax:   905-332-4844
Website:   www.aventix.ca
Email:   aventix@aventix.ca
Every effort has been made to ensure the accuracy of the ImmunoComb Feline VacciCheck information published above. However, it remains the responsibility of the readers to familiarize themselves with the product information contained on the US product label or package insert.

Copyright © 2019 Animalytix LLC. Updated: 2019-09-30

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