Shrimp Taura Syndrome Virus (TSV) Nucleic Acid Detection Kit (One Tube PCR-Fluorescence Probe Method) Instructions for Use

Shrimp Taura Syndrome Virus (TSV) Nucleic Acid Detection Kit (One-tube PCR-Fluorescence Probe Method)

â—† Product Description

The animal disease detection series can amplify specific nucleic acid fragments of pathogens in samples such as foods and animal tissues, and judge the results by real-time amplification curves. This product is used for the detection of Shrimp Taura Syndrome Virus (TSV) with a detection limit of 10 ³ copies/μL of genomic DNA .

• Product composition (96 test)

• Applicable instrument

Real-time fluorescence PCR instrument such as ABI 7500, CFX 96, Mx 3005P, LineGene9600.

â—† Self-supplied supplies and instruments

1 ice box; 2 pipettes (0.5-10μL, 10-100μL, 100-1000μL) and matching sterilization tips; 3 centrifuges; 4 vortex mixers; 5 metal baths; 6 homogenizers, mixers or Grinding tools such as mortar; 7 electronic balance.

â—† Notes

1. This reagent has high detection sensitivity. In order to prevent pollution, the experiment is to be partitioned.

1) First zone: sample preparation zone.

2) The second zone: the template addition zone.

3) Zone 3: Amplification and product analysis zone.

★ It is best to physically isolate the partitions to avoid contamination caused by human factors.

2. Work clothes and latex gloves are worn during the experiment, and tools are used independently in different areas. Gloves and lab coats need to be replaced.

3. Strictly follow the operation steps, reagent preparation and sample loading steps, please operate in strict accordance with the instructions on the ice box.

4. The components in the reaction solution are sensitive to light and should be stored away from light . The reagent should be completely thawed before use, but repeated freezing and thawing should be avoided. It is recommended to centrifuge for 30 seconds before use, and store the reaction solution in an appropriate volume according to the frequency of detection.

5. After the reaction is completed, the expansion tube should be placed in a sealed bag and discarded. On the same day, the lid is opened and the aerosol is easily contaminated. It is forbidden to open the lid.

6. Do not mix different batches of reagents used within the validity period.

â—† Sample processing

Samples prepared according to SN/T1151.1-2011 Shrimp Taura Syndrome Technical Specifications or other standards are prepared for use.

For detailed steps, please follow the standard operation or check the food safety software.

• Experimental operation

1. Template preparation (sample preparation area)

It is recommended to use the aquatic animal virus genomic DNA/RNA extraction kit series. The specific process is detailed in the product manual.

2. Add a template (template addition area, placed on the ice box)

Cut the PCR tube containing the reaction number, and place it at room temperature to be thawed. After centrifugation for 30 seconds, uncover the sealing film. Add 5 μL of template to each reaction solution in the order of NG, sample template to be tested. , PG-TSV-P. After the matching PCR tube cap was covered, the mixture was vortexed for 30 s, centrifuged for 1 min, and the PCR amplification reaction was immediately performed.

3. Amplification reaction (amplification and product analysis area)

Using a real-time PCR instrument, the fluorophore was selected for FAM and the quencher group was selected for TAMRA.

Set up the amplification reaction according to the following conditions:

For other instruments, please refer to the instrument manual for setting.

• Result determination

The test sample has no Ct value, the curve is straight or slightly oblique, there is no "S" type amplification curve, the sample can be reported negative, does not contain shrimp shrimp syndrome virus (TSV) or the content is below the detection limit;

The test sample has Ct≤38, and the curve shows an “S”-type amplification curve, which can directly report the sample positive, and contains the shrimp Taura Syndrome Virus (TSV);

Test sample 38

• The NG reaction is a smooth straight line, and the PG reaction is an "S" type amplification curve. The test result is valid, otherwise it is invalid. If the duplicate test results are still invalid, please contact technical support.
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