Step by Stepwith the

ImmunoComb®

The ImmunoComb®: A Diagnostic ToolThe ImmunoComb® is based on solid-phase ELISA principle. It provides serological results; namely, measuring antibody levels in blood or serum to specific disease agents (e.g., viruses, mycoplasma, rickettsia).

ImmunoComb® Antibody Test Kit - Contents

Developing Plate*Comb*

Tweezers*

Capillary Tube & piston

Comb Scale*

Note: * Included in all kits. Amount varies according to kit.

Specimen Paper*Specimen Paper*Specimen Paper*Specimen Paper*

Main Components

Antigens are attached to each tooth of the Comb. The placement of the antigens is shown by the labeling on the Comb.

The developing plate contains the reagents needed for Comb development.

Preparing ImmunoComb® Kit for Use

The ImmunoComb® kit is based on an enzymatic reaction, therefore it requires warm temperature. There are two options to reach proper temperature:

One option:

Allow to stand inroom temperature

of 20° – 25° C (68° – 77° F)

for 1 - 2 hours.

Another option:

Warm it in a 37° C/98.6° F incubator

for 20 minutes.

Obtaining Blood/Serum from Animal

1. Take a specimen paper with prepunched disks.

2. Saturate one pair thoroughly (both sides) with blood.

3. Allow to air dry.

A. When using specimen paper

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

A. When using specimen paper - Continued

5. Punch out one disk of the pair, saturated with blood and immerse it into well of row A.

6. Store the second saturated disk in a refrigerator for possible later use.

7. Wait 60 minutes before proceeding.

ATTENTION: Do not open any well that you do not intend to use.

Obtaining Blood/Serum from Animal

B. When using serum or whole blood

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

2. For testing whole blood use 10l. For testing serum use 5l.

3. To transfer blood/serum sample from test tube into a well in row A, use either capillary tube or pipette.

ATTENTION: Do not open any well that you do not intend to use.

Capillary tube with

piston

B. When using serum or whole blood - Continued4. Deposit a sample into well in row A. Raise and

lower the piston/pipette plunger several times to achieve mixing.

5. No need for an-hour wait.

Developing the Kit after Inserting Sample(s)

1. Remove the Comb from its protective envelope. For testing less than 12 samples, cut Comb in allocated notches for the number of tests required.

2. Insert the Comb into the open well(s) with the sample(s) in row A (printed side facing you) and incubate.

3. To improve mixing, gently jiggle the Comb up and down at the start of each incubation. Repeat this motion every 2-3 minutes for achieving best results.

Timing of incubation depends on type of kit.

Developing the Kit after Inserting Sample(s)

4. Transfer the Comb to the remaining wells (B-F) at timed intervals, according to instruction manual enclosed in kit.

5. Before moving the Comb from one well to the next, pierce with tweezers the foil of next well. Gently shake off excess liquids from Comb’s teeth on a clean paper towel.

6. When developing process is over, allow Comb to air dry completely.

Analyzing Results

Example of a Developed Comb for testing

Canine Parvovirus & Distemper (3 Teeth)

Positive ReferenceCPVCDV

The color intensity of spots corresponds to antibody level.

Analyzing Results

Example of a Developed Comb for testing

Poultry IBD – ND –IB

The color intensity of spots

corresponds to

antibody level.

1 2 3 4

IBD

ND

IB

C+

Positive control

C-

Negative

control

Analyzing Results

Example of a Developed Comb for testing

Bovine Leptospira

Internal Control

L. Hardjo

1 2 3

C+

Positive control

C-

Negative

control

The color intensity of

spots correspond

s to antibody level.

There are three ways to read results: (1) by eye, (2) by using the Comb Scale and (3) by using the CombScan 2000.

Reading results by eye

Provides qualitative results

Reading results using the Comb Scale

Provides semi-quantitative results

Reading results using the Comb Scan 2000

Provides quantitative results

Scoring Results

Reading Results by Eye

Compare the color tone of the test spot with either the positive reference spot (in pet kits) or with the positive control (livestock and poultry kits).

There are 4 levels of interpretation:

Negative

Low Positive (suspicious)

Positive

High PositiveCharts in manual instruction help user with interpretation of results.

1. Look for the color tone of the positive reference spot on the color scale of the Comb Scale.

2. Slide the yellow ruler until the C+ mark appears in the window corresponding to that color of the positive reference spot.

Reading Results using the CombScale

Continued – using

CombScale

3. Hold the ruler in this position while finding the matching color tone of each test spot.

4. The number that appears in the window above the color tone just found is the Comb Scale score (S0 – S6).

Reading Results using the CombScan 2000

The CombScan 2000 is a software program, which utilizes a computer and an office scanner.

When a Comb is placed on the scanner the

program translates the color results into numerical

values. It is free of charge product and it is most useful for large number of tests.

Summary

Contents of kit

Preparing the kit for use

Obtaining blood/serum

a. Using paper disks

b. Using capillary tubes or pipette

Developing the kit

Reading results and interpreting

a. By eye

b. Using the CombScale

c. Using the CombScan 2000

This is an introductory presentation. In order to

achieve best results using the ImmunoComb® test Kit you must read and follow

the instructions in the manual instruction provided

in each kit.

Thank You!

For Technical Support Please Contact:

Biogal – Galed Labs

Tel: 972-4-9898605, E-mail: info@biogal.co.il

www.biogal.co.il