SN/T 5107-2019 in English
VALIDA multiplex liquid bead array for detecting mosquito-borne diseases of multiplex pathogens at entry-exit ports
- Issued on:2019-09-03
- Implemented on:2020-03-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$146.00
Introduction
Analysis of Standard Technical Framework
| Detection object | Sample type | Sensitivity requirement | Detection time |
|---|---|---|---|
| Yellow Fever Virus | Serum/Mosquitoes | ≥10^3 copies | 6 hours |
| Dengue Virus Type Ⅰ-Ⅳ | Whole blood/Mosquitoes | ≥10^2 copies | 5.5 hours |
| Plasmodium | Anticoagulation | ≥1 parasite/μL | 7 hours |
Core Technology Principles
Gene suspension chip technology uses carboxyl-encoded microspheres (diameter 5.6μm) as probe carriers and adopts the principle of flow cytometry to achieve simultaneous detection of multiple targets. Key technological breakthroughs include:
- Multiplex PCR amplification: Set up 3 independent reaction systems to cover all pathogens
- Probe design: 20bp poly(T) spacer at the 5' end to improve hybridization efficiency
- Signal amplification: Streptavidin-phycoerythrin (SA-PE) secondary amplification system
Key points of biosafety management
The standard requires that the test must be carried out in a BSL-2 laboratory, with the following key control points:
- Sample pretreatment must be completed in a biosafety cabinet
- Used pipette tips should be soaked in 10% sodium hypochlorite solution
- PCR amplification products must be sterilized at 121°C under high pressure
Implementation and application suggestions
| Steps | Optimization measures | Quality control indicators |
|---|---|---|
| Nucleic acid extraction | Chelex-100 method was used to replace phenol-chloroform extraction for mosquito samples | A260/A280 ratio 1.8-2.0 |
| Microsphere coupling | EDC is prepared and used immediately (valid for ≤15 minutes) | Microsphere concentration ≥5000/μL |
| Hybridization reaction | Strict temperature control 55±0.5℃ | Blank control MFI value ≤200 |
Technology Evolution Analysis
Compared with the traditional PCR method, this standard technology has significant advantages:
- Detection throughput increased by 8 times (single detection of 6 types of pathogens)
- Cost reduced by 40% (saving on reagents and consumables)
- Objective interpretation of results (quantitative analysis of MFI values)
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