Step by Step

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Step by Step with the ImmunoComb
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The ImmunoComb A Diagnostic Tool
The 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).
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ImmunoComb Antibody Test Kit - Contents
Developing Plate
Comb
Specimen Paper
Specimen Paper
Specimen Paper
Specimen Paper
Tweezers
Comb Scale
Capillary Tube piston
Note Included in all kits. Amount varies
according to kit.
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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.
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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
Another option Warm it in a 37 C/98.6 F
incubator for 20 minutes.
One option Allow to stand in room
temperature of 20 25 C (68 77 F) for 1
- 2 hours.
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Obtaining Blood/Serum from Animal
A. When using specimen paper

• Take a specimen paper with prepunched disks.
• Saturate one pair thoroughly (both sides) with
  blood.
• Allow to air dry.

4. Use the tweezers to pierce protective
aluminum cover of row A. One well for each sample.
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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.
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Obtaining Blood/Serum from Animal
B. When using serum or whole blood

• Use the tweezers to pierce protective aluminum
  cover of well in row A. One well for each
  sample.
• For testing whole blood use 10?l. For
  testing serum use 5?l.
• To transfer blood/serum sample from test tube
  into a well in row A, use either
  capillary tube or pipette.

Capillary tube with piston
ATTENTION Do not open any well that you do not
intend to use.
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B. When using serum or whole blood - Continued
4. 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.
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Developing the Kit after Inserting Sample(s)

• Remove the Comb from its protective envelope.
  For testing less than 12 samples, cut Comb in
  allocated notches for the number of tests
  required.
• Insert the Comb into the open well(s) with the
  sample(s) in row A (printed side facing you) and
  incubate.
• 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.
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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 Combs teeth on a clean
paper towel. 6. When developing process is
over, allow Comb to air dry completely.
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Analyzing Results
Example of a Developed Comb for testing Canine
Parvovirus Distemper (3 Teeth)
Positive Reference CPV CDV
The color intensity of spots corresponds to
antibody level.
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Analyzing Results
Example of a Developed Comb for testing Poultry
IBD ND IB
C Positive control
IBD ND IB
The color intensity of spots corresponds to
antibody level.
C- Negative control
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Analyzing Results
Example of a Developed Comb for testing Bovine
Leptospira
C Positive control
Internal Control L. Hardjo
The color intensity of spots corresponds to
antibody level.
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C- Negative control
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Scoring Results
There are three ways to read results (1) by eye,
(2) by using the Comb Scale and (3) by using the
CombScan 2000.
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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 Positive
Charts in manual instruction help user with
interpretation of results.
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Reading Results using the CombScale

• Look for the color tone of the positive reference
  spot on the color scale of
  the Comb Scale.
• Slide the yellow ruler until the C mark appears
  in the window corresponding to that color of the
  positive reference spot.

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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).
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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.
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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
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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.
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Thank You!
For Technical Support Please Contact Biogal
Galed Labs Tel [phone], E-mail
info_at_biogal.co.il www.biogal.co.il