Infectious hypodermal and hematopoietic necrosis virus(IHHNV) Nucleic Acid Detection Kits

(PCR-Fluorescent Probe Method)

Intended use:

This kit is suitable for the specific detection of IHHNV in water samples, shrimp seedlings, bait, etc., and is used for the etiological diagnosis of suspected infected persons.

Principle:

This kit is chosen infectious subcutaneous and hematopoietic organ necrosis virus (IHHNV) specific fragment to design primers probe probe, the probe with primer amplification area in the middle of a specific binding DNA template, in the process of PCR extension, the circumscribed Taq polymerase enzymes from the 5 'end of fluorescent groups on the probe will be cut down, make it free in the reaction system, Thus, the 3 '-end quenched group was separated and fluorescence was emitted, which was detected by fluorescence PCR instrument to realize the detection of infectious subcutaneous and hematopoietic necrosis virus (IHHNV) in a completely closed reaction system.

Main components:

Sample requirements:

1. Sample type

Seedlings: parent, shrimp eggs, seedlings, coarse standard products

Pond: bottom mud, excrement, water body

Bait: live bait, chilled bait and non-high-temperature treated bait samples

2. Transport

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.