HiPer® Latex Agglutination Teaching Kit

Product Code: HTI001

Number of experiments that can be performed: 20

Duration of Experiment: ~ 15 minutes

Storage Instructions:

The kit is stable for 12 months from the date of receipt

Store the Antigen solution, Positive and Negative control at 2-8oC

Other kit contents can be stored at room temperature (15-25oC)

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 Index

Sr. No. Contents Page No.

1 Aim 3

2 Introduction 3

3 Principle 3

4 Kit Contents 4

5 Storage 4

6 Important Instructions 4

7 Procedure 4

8 Observation and Result 5

9 Interpretation 5

10 Troubleshooting Guide 5

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Aim:

To learn the technique of latex agglutination.

Introduction:

Agglutination is a reaction of clumping together of antigen-bearing cells, microorganisms or particles in the

presence of specific antibodies (agglutinins) in a suspension. Reaction time for agglutination to occur is
shorter compared to other antigen-antibody interactions. Latex agglutination makes use of latex particles
which are built from different organic materials to a desired diameter, and may be functionalized with
chemical groups to facilitate attachment of molecules. Latex agglutination tests have been in use since 1956
to detect a wide range of analytes in the clinical laboratory. The first description of a test based on latex
agglutination was the ‘Rheumatoid Factor Test’ proposed by Singer and Plotz in 1956. It can be used for
detection of both antigen and antibody.

Principle:

In latex agglutination, antibodies are adsorbed to the latex particles (under appropriate ionic and alkaline pH
conditions) by binding to the Fc region of antibodies leaving Fab region free to interact with antigen present in
the applied specimen. The use of smaller latex particle has improved the sensitivity and reagent longevity of
latex agglutination.

Antibody coated Visible Antigen coated Visible

latex particles clumping latex particles clumping

Applications of Latex Agglutination Tests:

Latex agglutination tests are very popular in clinical laboratories. These tests are applied to the detection of
many infectious diseases.

To detect microbial and viral infections, autoimmune diseases, hormones, drugs and serum
proteins.

To check for certain antibodies or antigens in a variety of bodily fluids including saliva, urine,
cerebrospinal fluid, and blood.

HiPer® Latex Agglutination Teaching Kit applies the above principle for rapid screening of multiple samples
for infection, microbial contamination or microbial identification. For ease of handling, the Latex Agglutination
Teaching Kit is provided with ready to use reagents. The kit gives direct results in form of visible
agglutination. In this kit three solutions are provided, i.e. Antigen solution, Negative control & Positive
control. The Antigen solution (preserved with 0.099% sodium azide) contains inactivated antigens reactive
with Test reagent and non-reactive with Control reagent. The Negative control contains latex particles coated

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with non-specific antibodies preserved in 0.099% sodium azide. The Positive control contains latex particles
coated with specific antibodies preserved in 0.099% sodium azide.

Kit Contents:

Table 1: Enlists the materials provided in this kit with their quantity and recommended storage

Sr. No. Product Quantity Storage

Materials Provided
Code 20 expts
1 TKC121 Antigen Solution- Solution A 1.08 ml 2-80C
2 TKC122 Negative Control- Solution B 0.54 ml 2-80C
3 TKC123 Positive Control- Solution C 0.54 ml 2-80C
4 TKC124 Disposable Agglutination Cards 7 Nos. RT
5 TKC125 Disposable Mixing Sticks 46 Nos. RT

Storage:

HiPer® Latex Agglutination Teaching Kit is stable for 12 months from the date of receipt without showing any
reduction in performance. On receipt, store the Antigen solution, Positive control and Negative control at 2-
8oC.

Important Instructions:

1. Latex Test Kit should be used according to the kit instructions.

2. Allow all reagents to reach room temperature before use.
3. Do not dilute any of the kit reagents.
4. Do not intermix the reagents.
5. Do not freeze any of the kit reagents.
6. Ensure the agglutination card is clean and dry prior to use.
7. Wear gloves while handling the reagents.

Procedure:

1. Before starting the experiment, gently mix all the bottles provided in this kit.
2. Add 1 drop of Solution A (25µL) into the circles marked as 1 and 2 of a clean dry agglutination card.
3. Add 1 drop of Solution B (25µL) into circle 1.
4. Add 1 drop of Solution C (25µL) to circle 2.
5. Spread the drops over the area of both the circles using fresh mixing stick for each circle.
6. Rock the card gently (approximately two to three minutes) and observe for agglutination. An
agglutination reaction is indicated by visible aggregation of the latex particles.
7. The circles marked as 3, 4 and 5, 6 can be used similarly
8. After performing the experiment, discard the slides and mixing sticks.

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Observation and Result:

After mixing the Antigen Solution with Positive control and Negative control separately observe for the
agglutination reaction.

1: Corresponds to Negative result

2: Corresponds to Positive result

Interpretation:

The results can be interpreted as follows:

Circle Number Test Interpretation

1 Negative No Agglutination seen
2 Positive Agglutination seen

• For circle 1, the inactivated antigens of the Antigen solution do not bind to the latex particles coated
with non-specific antibodies of the Negative control. Hence, no agglutination is seen.
• For circle 2, the inactivated antigens of the Antigen solution bind to the latex particles coated with
specific antibodies of the Positive control. Hence, agglutination is seen.

Troubleshooting Guide:

Sr.No Problem Possible Cause Solution

False positive The reagents were Ensure that the reagents are added properly onto
1 result mixed with each other the respective well without spilling to the sides
Incubated for a longer The results should be read within the time period
time mentioned in the brochure
No agglutination The reagents are not Ensure that the latex reagent, positive and
2 observed stored under proper negative control are stored in refrigerator
conditions (2-8oC)

Technical Assistance:
At HiMedia we pride ourselves on the quality and availability of our technical support. For any kind of
technical assistance mail at mb@himedialabs.com
PIHTI001_O/0514 HTI001-03