Vibrio parahaemolyticus kit Nucleic Acid PCR Detection Kit

(PCR-Fluorescent Probe Method)

Intended use:

This kit is suitable for the specific detection of pathogenic vibrio parahaemolyticus (Vpa) in bait, water samples, shrimp seedlings, shrimp tissues and processed products.

Principle:

This kit uses a specific fragment of Vibrio parahaemolyticus (Vpa) to design a primer probe probe, which can specifically bind to a SECTION of DNA template in the middle of the primer amplification region. During the PCR extension process, the exonuclide activity of Taq enzyme cuts the 5 '-terminal fluorophore from the probe and makes it free in the reaction system. Thus, the 3 '-end quenched group was separated, and the fluorescence was emitted, which was detected by fluorescence PCR instrument to achieve the detection of Vibrio parahaemolyticus nucleic acid (Vpa) in the completely closed reaction system.

Main components:

Sample requirements:

1. Sample type

Shrimp seedling, shrimp tissue, water body, bait, processed products, etc

2. Sample collection and transportation

Sample collection and transportation shall be carried out in accordance with NY/T 541. Samples shall be transported to the laboratory as soon as possible under refrigerated conditions to avoid repeated freeze-thaw.

3. Sample preservation

It can be stored for 24 hours at 2℃~8℃, and can be stored for a long time at -20℃.

Using Micgene 242/244/244IVD and ABI QuantStudio5:

procedures are set as below:

FAM was selected for detection channel.

The results of interpretation

Note: The result is judged to be grey area, which needs to be rechecked. If the recheck result is Ct value < 45 is considered as positive, and no Ct value or Ct value ≥45 is considered as negative.

Result analysis:
Automatic analysis using instrumentation

Quality control:
Negative control: No obvious amplification curve of FAM detection channel;
Positive control: FAM channel showed obvious amplification curve, Ct value ≤30;

The above conditions are met simultaneously in the same experiment; otherwise, the experiment is invalid and re-testing is required.