SN/T 5145.2-2019 in English
VALIDRapid Identification of animal ingredient in export food and feed Part 2: Deteetion of Martes Ingredient PCR- Lateral flow dipstick method
- Issued on:2019-10-25
- Implemented on:2020-05-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$117.00
Introduction
Technical background of the standard
As an important part of the import and export food safety testing series, this standard adopts the PCR lateral flow test strip technology to achieve rapid testing in response to the needs of animal-derived feed ingredient identification. It was issued on October 25, 2019 and officially implemented on May 1, 2020.
Core Detection Principle
| Technical Links | Key Technologies | Parameter Requirements |
|---|---|---|
| DNA Extraction | CTAB Method | Purity A260/A280=1.7-1.9 |
| PCR Amplification | Specific Primers | Annealing Temperature 55℃ |
| Test Strip Detection | FITC/Biotin Double Labeling | Detection Limit 0.1% |
Key Operation Points
DNA Extraction Process
Use CTAB lysis buffer (20g/L CTAB + 1.4mol/L NaCl) in a 65℃ water bath for 1 hour, extract with chloroform/isoamyl alcohol, and precipitate DNA with 70% ethanol. Pay special attention to the sample size must be ≥100mg to ensure extraction efficiency.
PCR system configuration
The standard reaction system includes:
- 2μL 10× PCR buffer (containing 20mM Mg2+)
- 0.8μL upstream and downstream primers (5μmol/L)
- 0.2μL ExTaq enzyme (5U/μL)
- 1μL DNA template (4ng/μL)
Result judgment criteria
The following conditions must be met at the same time:
1. Positive control: both the quality control line and the test line are colored
2. Negative control: only the quality control line is colored
3. Sample detection: two lines are colored and judged as positive, and the similarity with the NCBI database must be ≥97%
Technical advantage comparison
Compared with the traditional ELISA method, this standard PCR-lateral flow test strip technology has the following advantages:
- Detection time is shortened to 3 hours (originally 8 hours)
- Sensitivity is increased by 10 times (0.1% vs 1%)
- FITC and biotin dual-labeled products can be detected simultaneously
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