White Spot Syndrome Virus（WSSV）Nucleic Acid Test Kit (PCR-Fluorescent Probe Method) 24T

This product needs to be packed with ice bags and shipped through FedEx or UPS.
It will arrive within 7-10 days. The shipping fee is included in the product price.

Product introduction：

This test kit offers a rapid and accurate method for detecting White Spot Syndrome Virus (WSSV) in shrimp.

• Technology: Employs real-time fluorescent quantitative PCR (TaqMan probe method) for sensitive and specific detection.
• Targets: Utilizes specific primers and fluorescent probes designed to target the unique nucleic acid sequence of WSSV.
• Applications: Detects WSSV in various shrimp samples, including:
  • Animal tissues
  • Feed
  • Other sample types (consult kit specifications)
• Dual Detection: Provides qualitative and quantitative results:
  • Qualitative: Determines the presence or absence of WSSV.
  • Quantitative: Measures the amount of WSSV nucleic acid present, allowing for assessment of viral load.
• Real-Time Monitoring: Utilizes real-time PCR for continuous monitoring of amplification throughout the reaction, enabling rapid results.

This user-friendly test kit provides a valuable tool for shrimp farmers, researchers, and diagnostic labs for effective WSSV detection and management.

Product contents：

Storage:

Store at -20°C, with a shelf life of at least 12 months.

Applicable instrument：

ABI series (ABI 7300/ABI 7500/Step One, etc.), Roche LightCycler series, Bio-Rad CFX 96, SanshiBio Q162D, and other real-time fluorescence quantitative PCR instruments

Sample processing：

Process the samples according to relevant standards, and store the processed samples for later use.

Experimental procedures：

• Sample Preparation (Sample Preparation Area)

Please refer to the instruction manual of the total DNA/RNA extraction kit, or use the ES08 fully automated nucleic acid extraction and purification instrument with a matching rapid nucleic acid extraction kit (magnetic bead method), or other nucleic acid extraction kits/methods that comply with relevant standards to extract nucleic acid from the processed samples. The extracted sample nucleic acid should be tested as soon as possible and stored in an icebox, or else stored at -20°C.

• Reaction System Preparation (Adding Area)

2.1 Take out the WSSV positive control tube, negative control tube, and reaction tubes containing the WSSV reaction solution required for the experiment (n+2) (i.e., n samples to be tested + positive control + negative control), completely thaw the reagents at room temperature, centrifuge for 10 seconds, centrifuge the liquid from the tube wall and lid to the bottom of the tube, and store in an icebox for later use.

2.2 Then add 2μL of extracted negative control, sample nucleic acid, and WSSV positive control to the reaction tube in the eight-row strip, close the tube lid, make a record, and make the total volume of each reaction 25μL. Mix thoroughly, centrifuge for 30 seconds, and perform the amplification experiment on the real-time fluorescence quantitative PCR instrument.

• Fluorescent PCR Amplification (Amplification and Product Analysis Area)

Pre-denaturation at 95°C for 3 minutes; denaturation at 95°C for 5 seconds, annealing and extension at 60°C for 30 seconds, for 40 cycles, collect fluorescence signal at 60°C. Choose FAM as the fluorescence group, and None as the quencher group (for use with ABI series real-time fluorescence quantitative PCR instruments, if necessary, contact the manufacturer in advance or add ROX calibration dye by yourself, and select ROX as the quencher group; otherwise, follow the normal procedure).

• Result Determination

4.1 Ct value of the positive control < 30 and an “S” shaped amplification curve appears, the test result is valid. If not, the experiment should be repeated. If the retest is still invalid, please contact the technical personnel.

4.2 Sample detection result: Ct value ≤ 38 and an “S” shaped amplification curve appears, indicating a positive result, which means the sample contains White Spot Syndrome Virus (WSSV).

4.3 Sample detection result: 38 < Ct < 40, it is considered suspicious. The sample should be retested. If the retest Ct value < 40 and there is a clear amplification curve, it is considered positive; otherwise, it is considered negative.

4.4 Sample detection result: No Ct value or Ct value ≥ 40 and no “S” shaped amplification curve, it is considered negative, indicating that White Spot Syndrome Virus (WSSV) was not detected in the sample.

Precautions：

• This test kit has high sensitivity. To prevent contamination, the experiment needs to be strictly partitioned, and physical isolation between partitions is recommended to avoid cross-contamination caused by human factors.
• Wear lab coats and latex gloves during the experiment. Use separate tools in different areas, and remember to change gloves and lab coats.
• The components in the reaction solution are light-sensitive and should be stored away from light. Thaw the reagents completely before use, but avoid repeated freezing and thawing.
• Follow the instructions strictly for steps such as reagent preparation and sample addition. Disinfect the workbench, centrifuge, and pipettor regularly using chlorine disinfectants, ethanol, nucleic acid decontamination agents, or ultraviolet light.
• After the reaction, place the PCR reaction tubes in sealed bags for disposal. Do not open the tubes to avoid aerosol contamination.
• Do not mix reagents from different batches. Use within the expiration date. Consumables should be treated without enzymes and sterile.
• A negative test result does not completely indicate a non-infected state; it is closely related to the quality of the obtained nucleic acid. “Negative” results may be due to unqualified nucleic acid sample quality, low sample nucleic acid load, or unsuccessful nucleic acid extraction (detection). Veterinary clinical considerations are necessary for specific diagnoses.
• Possible causes of false positives: Cross-contamination during sample collection, transportation, and nucleic acid extraction.
• For any other questions, please contact the manufacturer’s technical staff promptly. This product is for scientific research use only and is not intended for clinical diagnosis or other purposes.