SC/T 7204.5-2020 in English
VALIDDiagnostic protocols for taura syndrome of penaeid shrimp-Part 5: Reverse transcription loop-mediated isothermal amplifcation
- Issued on:2020-08-26
- Implemented on:2021-01-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$127.00
| Standard No: | SC/T 7204.5-2020 |
| Document status: | VALID |
| Title in English: | Diagnostic protocols for taura syndrome of penaeid shrimp-Part 5: Reverse transcription loop-mediated isothermal amplifcation |
| Title in Chinese: | 对虾桃拉综合征诊断规程第5部分：逆转录环介导核酸等温扩增检测法 |
| Language: | English |
| File Format: | Electronic (PDF) |
| Delivery: | Via email within 1~3 business days |
| Issued on: | 2020-08-26 |
| Implemented on: | 2021-01-01 |
| Professional Classification: | SC-Aquaculture |
| Related Keywords: | taura syndrome
loop-mediated isothermal amplifcation introduction standard technical background sc/t area amplification area result interpretation area environmental treatment reverse transcription diagnostic protocols |
| Related Topics: | mediate
amplify reverse transcription sc test Reverse transcription at 70 degrees for 5 minutes loop amplification isothermal amplification Amplification detection "Reverse Transcription Qitian isothermal amplification of COVID-19 Transcription-mediated amplification technology |
Introduction
Standard Technical Background
SC/T 7204.5-2020, as the fifth part of the series of standards for the diagnosis of Taura syndrome in shrimp, for the first time incorporates the RT-LAMP technology into the aquaculture industry standard system. This technology can complete TSV RNA detection within 60 minutes through 63℃ constant temperature amplification, which is more than 3 times more efficient than traditional RT-PCR.
Core Detection Process
| Steps | Key Parameters | Technical Points |
|---|---|---|
| RNA Extraction | 10-100 ng/μL | Denaturation at 65℃ for 15 minutes |
| Reaction System | 24μL/tube | Contains 1.2 mol/L betaine |
| Amplification Conditions | 63℃ 60min | SYBR Fluorescence Channel Monitoring |
Technical Advantages Analysis
The TSV-FIP/BIP primers designed in the standard (see Appendix B) target sites 56-66 of the GenBank AY997025 sequence, forming a stem-loop structure to achieve exponential amplification. Compared with traditional methods:
- Sensitivity reaches 102 copies/μL
- No thermal cycler required
- Visual interpretation of "S" curve
Laboratory Management Standards
Appendix C clearly requires physical isolation of four areas:
- Clean area (reagent preparation)
- Sample processing area
- Amplification area
- Result interpretation area
Implementation suggestions
1. Store primers in aliquots to avoid repeated freezing and thawing
2. Establish a quality control system for positive standards (ATCC VR-1392)
3. Perform method validation in accordance with SC/T 7204.3
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