WARNINGS AND PRECAUTIONS

WHITE SPOT SYNDROME VIRUS (WSSV) RAPID TEST KIT

INTENDED USE
White Spot Syndrome Virus (WSSV) Rapid Test Kit is an immunochromatographic assay designed for the
qualitative detection of WSSV infection in shrimp.
SUMMARY AND EXPLANATION OF TEST
WSSV is a highly infectious agent that causes mass mortality in farmed penaeid shrimp worldwide. Detection
of WSSV in shrimps is very important for management of shrimp farms. WSSV Rapid Test kit is a
management tool based on immunoassay for shrimp aquaculture that can be used to assess the level of
WSSV infection in shrimp. The beneficial features of this kit are that simple, convenient and quick results can
be obtained without the requirement of sophisticated tools or special skills.
PRINCIPLE OF TEST
The test device consists of a membrane strip and plastic case. A colored monoclonal rabbit anti-WSSV
antibody colloid gold conjugate (antibody -colloid gold conjugate) pad is placed next to the sample pad region
on the membrane strip. The membrane strip is also pre-coated with different monoclonal rabbit anti-WSSV
antibody on the test (T) zone and goat anti-mouse ImmnoglobulinG (IgG) antibody on the control (C) zone.
In the absence of WSSV in the shrimp, the antibody-colloid gold conjugate moves with the sample by
capillary action along the membrane through the T zone until it reaches the C zone. At this point, antibody-
colloid gold conjugate reacts with the pre-coated goat anti-mouse IgG antibody to produce a visible pink
colored band. When only one pink colored band appears on the C zone, it means negative. When the WSSV
is present in the shrimp, the antibody-colloid gold conjugate reacts with WSSV to produce conjugate-WSSV
complex and moves until it reaches the T zone. At this point, the complex reacts with the pre-coated different
monoclonal rabbit anti-WSSV antibody to produce a visible pink colored band. An excess free antibody-
colloid gold conjugate passes through the T zone until the C zone. At this point, the excess antibody-colloid
gold conjugate reacts with the pre-coated goat anti-mouse IgG antibody to produce a visible pink colored
band. When the pink colored band appears in both T zone and C zone, it means positive.
REAGENTS AND MATERIALS SUPPLIED MATERIALS REQUIRED BUT NOT SUPPLIED
1. For detecting WSSV in the shrimp only.
2. The buffer liquid in the dropper bottle should be used for virus testing purpose only. Avoid contact
with skin or eyes.
3. This product insert must be strictly followed in order to produce accurate test results.
4. Keep the test device sealed until use. Once the device pouch has been opened, the test device must
be used immediately.
5. All test devices, reagents and specimens must be at room temperature (15-30°C) before running the
assay.
6. Do not use device if the sealed pouch is visibly damaged.
7. Do not use the kit contents beyond the expiration date.
8. Handle all specimens as being potentially infectious. Dispose all materials that come in contact with
the specimen as infectious waste.
9. Do not reuse test device.
10. Discard the used kit in a safe place.
11. The kit should be used immediately after open the package. Prolong expose the kit to the open air
deteriorate the kit and fault negative may occur.
12. If possible, wear gloves or wash hands with 70% alcohol before performing the test.
13. The used kit cannot be reused, it should be disinfected with 70% alcohol or bleach before discarded.
SPECIMEN COLLECTION AND PREPARATION
1. No special pre-treatment is necessary.
2. The specimen collection may be stored up to 2 days at refrigerated condition (2-8°C).
3. If it is necessary to store the specimen more than 3 days, it should be stored under frozen
condition (-20 ℃ or below).
QUALITY CONTROL
1. Positive and negative controls are not included and are optional.
2. If the control line at position C does not become visible, the test is invalid and the test must be
repeated. Positive samples will have additional coloured band at position T.

1. WSSV Rapid Test cassette (20 pieces) 1. Clock or timer ASSAY PROCEDURE
2. Buffer (2 dropper bottles) 2. Scissor
3. Microcentrifuge tube (20 pieces, single use only) 1. Bring test cassette and buffer to room temperature (15-30oC).
4. Disposable pestle (20 pieces, single use only) 2. Add 20 drops (approximately 0.5 ml) of buffer into the microcentrifuge tube provided with the kit.
5. Disposable dropper ( 20 pieces, single use only) 3. Put a piece of shrimp tissue sample, such as 1 gill filament, 1 swimmeret, or walking leg into the
6. One copy of instruction manual (product insert) microcentrifuge tube containing 0.5 ml of buffer.
4. Ground the sample into small pieces with the disposable pestle.
5. Gently tear open pouch and remove the test cassette. Lay the test device on a clean, flat work
surface.
STORAGE AND STABILITY 6. Using the disposable dropper/ transfer pipette, slowly apply 5- 6 drops (approximately 100-150 µl) of
The kit can be stored at room temperature in the sealed pouch to the date of expiration. The kit should be the sample solution into the application chamber/ sample well of the test cassette.
kept away from direct sunlight, moisture and heat. Expiration date is described on the product package. 7. Read the test result within 15 minutes.

Do not reuse same microcentrifuge tube or disposable pestle for different sample to avoid cross
contamination. Do not reuse the test cassette.
Do not observe the result after 15 minutes because after the strip is dried a light gray band of
hemocyanin deposition may be observed.

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DIAGRAM OF ASSAY PROCEDURE WARRANTY AND LIMITED LIABILITY
The performance characteristics stated were obtained by using the assay procedure in this insert. Failure to
follow the assay procedure may derive inaccurate results. In such event, the manufacturer disclaims all
warranties expressed, implied or statutory, including the implied warranty of merchantability and the fitness
for use. The manufacturer will not be liable for any damage caused by misuse, improper handling and
storage, non-compliance with warnings and procedures, damage caused by events occurring after the
product is released, failure to ensure the product is in proper condition before use, or any warranty given by
independent distributor.

REFERENCES

1. Sithigorngul W, Rukpratanporn S, Pecharaburanin N, Longyant S, Chaivisuthangkura P, Sithigorngul P (2006)

A simple and rapid immunochromatographic test strip for detection of white spot syndrome virus (WSSV) of
shrimp. Dis Aquat Org 72: 101–106
2. Sithigorngul P, Rukpratanporn S, Chaivisuthangkura P, Sridulyakul P, Longyant S (2011) Simultaneous and
rapid detection of white spot syndrome virus and yellow head virus infection in shrimp with a dual
immunochromatographic strip test. J Virol Methods 173:85–91
3. Sithigorngul P, Chaivisuthangkura P, Longyant S (2011) Immunological-based assays for specific detection of
shrimp viruses. World J Virol 3(1): 1-10

INTERPRETATION OF RESULT

ORDER INFORMATION

Product Code Description Packing Size

Positive result: RWSS01-020 White Spot Syndrome Virus Rapid Test Kit 20 tests/kit
The pink purple band should be observed at both the test line (T) and control line (C) indicating that the
shrimp was infected with WSSV.
MANUFACTURER DISTRIBUTOR
Negative result: Reszon Diagnostics International Sdn. Bhd. Profound Kestrel Laboratories Sdn Bhd
The pink purple band appear only on the control line (C) indicating that no infection of WSSV or light infection Revongen Corporation Center, Revongen Corporation Center,
beyond the sensitivity of the kit. The sample should be confirm by PCR. No.12A, Jalan TP5, Taman Perindustrian UEP, No.12A, Jalan TP5, Taman Perindustrian UEP,
47600 Subang Jaya, Selangor, Malaysia 47600 Subang Jaya, Selangor, Malaysia
Invalid: Tel : 603-8025 1603 Tel : 603-8025 1603
If there is no line at the control line(C), please retest with new kit Fax : 603-8025 1637 Fax : 603-8025 1637
Email : info@reszonics.com Email : lab@profoundkestrel.com
REMARK Website : www.reszonics.com Website : www.profoundkestrel.com
This kit is used for primary screening for WSSV infection. The negative result sample should be
confirmed by nested PCR or higher sensitive methods.

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