SN/T 5145.1-2019 in English

Technical background and evolution of the standard

As an important part of the entry-exit inspection and quarantine industry standard system, this standard, in response to the increasingly prominent problem of food adulteration in international trade, establishes for the first time a standardized process for the detection of cat-derived ingredients by the PCR-test strip method. Compared with the traditional electrophoresis detection method, the test strip technology shortens the detection time from 4 hours to 90 minutes and increases the sensitivity by 10 times.

Principle of the core method

Key Operation Points

1. Primer design: Use FITC-labeled forward primer (5'-CCATACATCGGGACTGAAC-3') and biotin-labeled reverse primer, and the amplification product is a 148bp fragment
2. DNA purity control: The A260/A280 ratio must be strictly controlled between 1.7-1.9, and re-purification is required if it exceeds the range
3. Temperature parameters: During DNA extraction, the 65℃ water bath must be accurate to ±1℃, and the PCR annealing temperature of 55℃ cannot be adjusted

Implementation suggestions

The laboratory should establish the following quality control system:

• Each batch of testing must include negative/positive controls and blank controls
• The test strips must be stored at 4℃ away from light. The validity period is shortened to 7 days after opening.
• Suspected positive results must be verified by sequencing (alignment with NCBI U20753.1 sequence ≥97%)

Technical Advantages Comparison