Transcript by using the Comb Scale and

Step by Step
with the
ImmunoComb®
The ImmunoComb®: A Diagnostic Tool
The ImmunoComb®
is based on solidphase 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
Comb*
Developing
Plate*
Specimen
Paper*
Tweezers*
Comb Scale*
Capillary
Tube & piston
Note: * Included in all kits. Amount varies
according to kit.
Main Components
The developing plate
contains the reagents
needed for Comb
development.
Antigens are attached
to each tooth of the
Comb. The placement
of the antigens is
shown by the labeling
on the Comb.
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:
Another option:
Allow to stand in
room temperature
of 20° – 25° C
(68° – 77° F)
for 1 - 2 hours.
Warm it in
a 37° C/98.6° F incubator
for 20 minutes.
Obtaining Blood/Serum from Animal
A. When using specimen paper
1. Take a specimen paper
with prepunched disks.
2. Saturate one pair
thoroughly (both sides)
with blood.
3. Allow to air dry.
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 10l.
For testing serum use 5l.
3. 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.
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.
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 Reference
CPV
CDV
The color intensity
of spots
corresponds to
antibody level.
Analyzing Results
Example of a Developed Comb for testing
Poultry IBD – ND –IB
C+
IBD
ND
IB
1
Positive
control
2
3
4
CNegative
control
The color
intensity of
spots
corresponds
to antibody
level.
Analyzing Results
Example of a Developed Comb for testing
Bovine Leptospira
Internal Control
Positive
control
L. Hardjo
1
2
C+
3
CNegative
control
The color
intensity of
spots
corresponds
to antibody
level.
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.
Reading results
by eye
Provides
qualitative results
Reading results using Provides
the Comb Scale
semi-quantitative results
Reading results using Provides
the Comb Scan 2000 quantitative 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 Positive
Charts in manual instruction help user
with interpretation of results.
Reading Results using the CombScale
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.
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: [email protected]
www.biogal.co.il