SN/T 5279-2020 in English
VALIDQuarantine protocol for infection with Herpesvirus anguillae
- Issued on:2020-12-30
- Implemented on:2021-07-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$175.00
Introduction
Core Interpretation of Standard Technical Specifications
SN/T 5279-2020 standard systematically regulates the molecular detection technology system of eel herpesvirus (HVA). Its technical points are mainly reflected in:
| Detection method | Target fragment | Sensitivity | Judgment standard |
|---|---|---|---|
| Conventional PCR | 394bp | 95% homology | Gel electrophoresis + sequencing verification |
| Fluorescent probe PCR | Cap protein gene | Ct value≤35 | Amplification curve analysis |
Key technical implementation points
1. Sample processing specifications
The standard clearly requires: Individuals with body length > 4cm need to have gill/liver/spleen/kidney tissues taken, and individuals ≤4cm can be tested as a whole fish. Recommended DNA extraction:
- Lysis buffer (10mM Tris-Cl+0.5% SDS)
- Proteinase K digestion for 1-2 hours (55℃)
- Phenol-chloroform extraction followed by ethanol precipitation
2. Detection quality control
A three-level control system must be established:
- Positive control (GenBank KX027736.1)
- Negative control (confirmed non-infected sample)
- Blank control (sterile water)
Standard evolution and technological innovation
Compared with the early diagnosis method that relied on clinical symptoms, this standard has three major breakthroughs:
Case: In 2021, Shenzhen Customs adopted this standard to test imported eel fry and found a suspected sample with a Ct value of 37. After repeated testing, it was confirmed to be positive, avoiding the risk of virus introduction.
Technical Comparison
| Version | Detection Limit | Time Consumption | Specificity |
|---|---|---|---|
| Traditional Pathology | 10^3 copies/μL | 3-5 days | 60-70% |
| SN/T 5279-2020 | 10^1 copies/μL | 4 hours | 99% |
Implementation Recommendations
1. Laboratory Configuration Requirements
- Must be equipped with a fluorescence PCR instrument and an electrophoresis system
- It is recommended to use a commercial DNA extraction kit (such as Qiagen DNeasy)
2. Cross-validation of results
For samples with fluorescence PCR Ct values of 35-38, the following should be done:
- Repeat the test twice
- Conventional PCR sequencing verification
- Combined with epidemiological surveys
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