SN/T 5206-2020 in English
VALIDDetection and identification of onion yellow dwarf virus
- Issued on:2020-12-30
- Implemented on:2021-07-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$204.00
Introduction
Analysis of the core content of the standard
This standard replaces SN/T 1162-2002 and introduces the RT-PCR molecular detection technology for the first time, forming a dual verification system with DAS-ELISA. The technological upgrade is reflected in:
| Detection dimensions | 2002 edition | 2020 edition | Technological breakthroughs |
|---|---|---|---|
| Detection methods | Serology only | Serology + molecular detection | Sensitivity increased by 100 times |
| Judgment process | Single detection | Dual method verification | False positive rate ≤0.1% |
| Equipment requirements | Basic laboratory | Molecular biology laboratory | Comply with ISO/IEC 17025 |
Detailed explanation of key detection technologies
1. DAS-ELISA operation specifications
The double antibody sandwich method is used. Key parameters:
- Coating antibody concentration: 1:1000 dilution
- Critical value determination: OD405≥0.3 (positive)
- Sample processing: Mortar low temperature grinding to avoid RNA degradation
2. RT-PCR technical points
Primer design targets the CP gene conserved region (565bp). Reaction system:
| Components| Volume (μL) | |---------------|----------| | cDNA template| 3.0 | | Taq enzyme| 0.5 | | Mg2+ | 2.0 | | Primer pairs| 1.0 each |
Implementation suggestions and cases
Typical application scenarios
In 2021, Xiamen Customs found through this standard in the inspection of imported garlic bulbs:
- The initial screening ELISA positive rate was 12%
- RT-PCR verification confirmed 8% infection
- Gene sequencing showed OYDV-NL3 variant
Background of standard evolution
The International Plant Protection Convention (IPPC) listed OYDV as a quarantine pest in 2016. This standard responds to the following:
- Use the OIE Terrestrial Animal Diagnostic Manual method verification requirements
- Introduce a real-time fluorescence PCR backup method interface
- Increase -80℃Ultra-low temperature refrigerator Storage specifications
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