ImmunoComb-FIP (Canada)
This page contains information on ImmunoComb-FIP for veterinary use.The information provided typically includes the following:
- ImmunoComb-FIP Indications
- Warnings and cautions for ImmunoComb-FIP
- Direction and dosage information for ImmunoComb-FIP
Immunocomb-fip
This treatment applies to the following species:A solid phase immunoassay for:
Feline Infectious Peritonitis Virus Antibody Test Kit
Instruction Manual
February 04, 2009
For IN VITRO use only
I. Intended Use Of The Kit
This kit is designed to aid in the diagnosis of Feline Infectious Peritonitis (FIP). A negative result is helpful in ruling out a diagnosis of FIP.
Ii. General Information
It is estimated that up to 70% of cats, worldwide, are exposed to Feline Enteric Corona Viruses (FECV). Infection is transmitted by the fecal-oral route; the virus can survive in dried secretions for as long as seven weeks. The risk of exposure is higher in catteries and multiple-cat households.
FECV infection in most cats is not associated with clinically apparent disease. In some cats, however, a severe, typically fatal, disease (known as FIP) may develop. FIP is believed to be caused by a mutant form of FECV, referred to as FIP virus (FIPV).
Iii. Clinical Signs
In most cats, infection with FECV is asymptomatic. In a small percentage of cases, fever, diarrhea and upper respiratory signs such as conjunctivitis can occur. This stage may last for an undefined time and then progress to a severe systemic disease known as Feline Infectious Peritonitis (FIP). FIP manifests clinically in 2 forms: effusive (wet) and non-effusive (dry). FIP is generally associated with a fatal outcome, even with therapy.
Iv. Diagnosis
Antibodies to FIPV indicate previous exposure to FECV. It is unclear why clinical disease (FIP) develops only in a small percentage of infected cats. Many of them have a history of recent stress, such as relocation to a new home, surgery (e.g., neutering) or another illness. Cats with FIP typically have antibodies to FIPV. As such, serology is considered to be useful for helping diagnose individual clinical cases as well as for prevention and control programs in multiple cat households or facilities. Yet a definitive clinical diagnosis should not be based entirely on the serological results.
V. What Is The Immunocomb®-fip Assay?
The ImmunoComb®-FIP test is a modified ELISA, which has been described as a “dot”-ELISA that detects antibody levels in serum or whole blood. The kit contains all necessary reagents for developing the test and is a self-contained portable kit. Results for the FIPV tests are obtained in less than 38 minutes.
Vi. How Does The Immunocomb®-fip Work?
- The ImmunoComb®-FIP kit contains 2 main components: comb shaped plastic card, hereafter referred to as the Comb and 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 spots of FIPV antigen are attached to the lowest spot on each tooth of the Comb. The middle spot is the Positive Reference and the upper most spot is the Internal Control (See Fig. 1).
- IgG antibodies to FIPV if present in serum, plasma or blood specimen used in row A of the multicompartment developing plate, react with the FIPV antigen on Comb test spots.
- At the end of the developing process, described in section VIII, if positive, a purple-grey color result develops via an enzymatic reaction.
- On each tooth of the developed Comb you should see the Internal Control spot (upper spot) and the Positive Reference spot (middle spot). The test spot of FIPV (bottom spot) may appear, depending on the result.
- Results are scored using the Positive Reference spot and the CombScale (scale S0-S6; see section X).
Vii. Kit Contents
1. The Immunocomb®-fip Card - One Comb Sufficient For 12 Tests.
fig 1: Two Teeth Of A Developed Comb (example)
2. Developing Plate (fig. 2) - One Plate, Divided Into Compartments A-f, That Are Subdivided Into 12 Wells; The Plate Compartments Are Pre-filled With The Reagent Solutions.
fig 2
3. Disposables Tweezers - For piercing the foil cover of developing plate compartments.
4. CombScale - One calibrated CombScale color card for scoring reactions intensities.
5. Heparinized Capillary Tubes - A vial with thirteen 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), 10 microliters blood (upper band).
6. Documentation - Instruction manual with detailed instructions.
VIII. STEP BY STEP WITH THE IMMUNOCOMB®-FIP
Perform Assay At Room Temperature Of 20° - 25° C (68° - 77° F).
(1) Obtain blood sample from cat.
(2) Use a pipette or a capillary tube*. For testing whole blood use 10 µl. For testing serum/plasma use 5 µl.
(3) Use the tweezers to pierce the protective aluminum cover of row A**. One well for each sample/specimen.
(4) Deposit a sample into a well in row A*. Raise and lower the pipette/piston plunger several times to achieve mixing.
Do Not Open Any Wells Of Row A Or Other Rows Which You Do Not Intend To Use.
(5) Remove the Comb from its protective envelope. For testing less than 12 samples, cut or fold Comb in allocated notches for the number of tests required. Insert the Comb into the open well(s) in row A** (printed side facing you) and incubate for 10 minutes. To improve mixing, gently dip the Comb up and down at the start of each incubation (each row). Repeat this motion every 2-3 minutes in all rows for achieving best results.
(6) Use tweezers to pierce the foil of the next well (row B**), and insert Comb for 2 minutes. Before transferring Comb from one well to the next, pierce the foil of the next well. Gently shake off excess liquid from Comb teeth onto a tissue and insert Comb into the next well (row C**) for 10 minutes. Then, place Comb in the remaining wells (row D & E**) for 2 minutes and the last well (row F*) for 10 minutes.
(7) Upon completion of 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 1-10 minutes.
* Unit of 40 capillary tubes & one piston may be purchased upon request.
** See Fig. 2.
Ix. Reading And Interpreting The Fipv Antibody Results
- The upper spot is the Internal Control - it should give a dark purple-grey color.
- The middle spot, the Positive Reference spot, should give a distinct purple-grey color. This is the same color tone that is generated by a significant positive response. This spot should be read as S3 on the CombScale (a scale of S0 to S6).
- The bottom spot on the Comb tests for FIPV.
- Compare the color tone of the FIPV spot (bottom one) with the Positive Reference spot (middle one). A clear, visible purple-grey dot indicates a positive response to FIPV, so does any result darker than the Positive Reference. Color fainter than the Positive Reference indicates a low response to FIPV.
- To evaluate the score, use the CombScale provided in the kit (see section X).
- Cats with FIPV usually have high antibody levels.
- A negative result (less than S1) indicates that the cat has not been exposed or had cleared the virus, and is free of FIPV.
X. Reading Results With The Combscale
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 (middle spot). Slide the yellow ruler until the C+ mark appears in the window above that color you just found.
Hold The Slide 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.
while Holding The Slide, Find The Tone Of Purple-grey On The Combscale That Most Closely Matches The test Result Spot (bottom Spot). The Number That Appears In The Window Above Is The Combscale Score (s0-s6). Repeat This Step With Every Test Spot Separately.
xi. Example Of A Developed Comb
Tooth No. |
Result |
Remarks (in Cats With Clinical Signs) |
|
1, 12 |
S0 |
No reaction to FIPV antigen; FIP negative |
|
7 |
≤ S1 |
Low, non specific reaction; considered negative. |
|
3, 5 |
S2 |
Suspicious reaction (retest recommended); FIP unlikely. |
|
6, 8 |
≥ S3 |
Positive reaction; FIP possible. |
|
9 |
Invalid |
High background colour - Invalid test. |
|
11 |
≥ S3 |
High background colour with positive reaction; FIP possible. |
|
2, 4 |
≥ S5 |
Strong positive reaction; FIP possible. |
|
10 |
Invalid |
No internal control and no positive reference - Invalid test. |
Another way to read the results is by using the CombScan. 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®-FIP results and conserving the data, and is supplied free of charge upon request.
As With All Diagnostic Tests, A Definitive Clinical Diagnosis Should Not Be Based Entirely On The Serological Results, But Should Only Be Made By The Veterinarian, After All Clinical And Laboratory Findings Have Been Evaluated.
Xii. Limitations Of The Kit
- As detectable levels of serum antibodies require 2-3 weeks to develop, a false negative result is most likely to occur when cats are tested shortly after the onset of infection.
- A false positive result occurs when serum antibodies bind non-specifically. This may be caused by a poor quality specimen (i.e. one that is collected and stored improperly) or by antibodies that react with antigen constituents other than those of the virus. False positive results are suspected when positive reactions are unexpected or cannot be reproduced.
- This kit has not been shown to distinguish antibodies against FIPV from antibodies against FECV.
- Any result >S1 but <S3 is considered suspicious. It is strongly recommended to customers to retest, by an alternative method, samples that give suspicious reactions, invalid reactions or unexpected positive results.
|
Study ID |
Sample Size |
Sample Type |
Sensitivity & Specificity, 95% Confidence Limit (CL) |
Positive and Negative Predictive Value, Kappa Statistic, Odds Ratio |
|||
|
+/+ |
-/+ |
+/- |
-/- |
||||
|
Ib |
84 |
6 |
1 |
99 |
Serum |
Sen: 93% [95% CL 86%, 97%] Spec: 99% [95% CL 95%, 100%] |
Positive Predictive Value = 99% Negative Predictive Value = 94% Kappa Statistic = 0.926 Odds ratio = 1385.9 |
a. Kit results/reference test results.
b. Data from a representative study.
I. ImmunoComb® FIP kit results compared to results obtained using an immunofluorescence assay (IFA).
Xiii. Storage & Handling
1. Store the kit under normal refrigeration (2° - 8° C or 36° - 46° F). Do not freeze the kit.
2. Before conducting the test, maintain all kit elements and specimens at room temperature - preferably for 60 - 120 minutes (or 22 minutes at 37° C or 98.6° F). Perform assay at room temperature of 20° - 25°C or 68° - 77° F).
3. Avoid spillage and cross-contamination of solutions.
4. Mix reagents by inverting developing plate several times prior to use.
5. Do Not Mix Reagents From Different Kits Or From Different Compartments Of The Same Kit.
6. Do Not Touch Teeth Of The Immunocomb®-fip Card.
7. When using developing plate, pierce the cover of each compartment according to the test procedure instructions. Do not remove the aluminum cover of developing plate all at once.
8. The ImmunoComb®-FIP Kit contains inactivated biological material. The kit must be handled and disposed of in accordance with accepted sanitary requirements. It is recommended to incinerate the kit after use.
Xiv. References
Addie, D. D. (1998). The diagnosis and prevention of FIP and recent research into feline Coronavirus shedding. ESVIM Proceedings: 8th Annual Congress of the European Society of Veterinary Internal Medicine.
Addie, D. D. (2000). Guest editorial: Clustering of feline Coronaviruses in multicat households. The Veterinary Journal, 159, 8-9.
Addie, D. D., et al. (2002). Evaluation of the feline Coronavirus antibody ImmunoComb®. 2nd International FCoV/FIP Symposium, Glasgow, UK.
Kiss, I., et al. (2000). Prevalence and genetic pattern of feline Coronavirus in urban cat populations. The Veterinary Journal, 159, 64-70.
Addie D. D. et al. (2004) Evaluation of an in-practice test for feline coronavirus antibodies. Journal of Feline Medicine and Surgery, 6, 63-67.
For Further Assistance Please Contact
Modern Veterinary Therapeutics, LLC Coral Gables, Florida 33146 - USA
Tel: +1 305 669 4150, Fax: +1 305 232 6645
E-mail: info@modernveterinarytherapeutics.com
Site: www.modernveterinarytherapeutics.com
or: Biogal, Galed Labs., Galed, 19240, Israel Tel: 972-4-9898605, Fax: 972-4-9898690, E-mail: info@biogal.co.il
Site: www.biogal.co.il
Distributed In Canada By
Biovet-inc. - 4375 Avenue Beaudry, St-Hyacinthe, QC - Canada J2S 8W2Tel: +1 888 824-6838, Fax: +1 (450) 771-4158, Site: www.biovet-inc.com
Product made in Israel
Produced by: Biogal, Galed Labs., 19240, Israel
Produced for: Modern Veterinary Therapeutics, LLC
Distributed by: Biovet-inc. - St-Hyacinthe, QC Canada J2S 8W2
U.S. Permit No. 544-A
Instruction Cat. No: 63FIP420
Presentation
12 tests per kit.Nac No.
13540000 |
Every effort has been made to ensure the accuracy of the ImmunoComb-FIP information published above. However, it remains the responsibility of the readers to familiarize themselves with the product information contained on the Canadian product label or package insert. |
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