SN/T 2101-2016 in English

VALID

Determination of Mycobacterium tuberculosis in milk and dairy products for export - Real-time PCR method

Issued on:2016-03-09
Implemented on:2016-10-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $210.00
$204.00

Standard No:	SN/T 2101-2016
Document status:	VALID
Title in English:	Determination of Mycobacterium tuberculosis in milk and dairy products for export - Real-time PCR method
Title in Chinese:	出口乳及乳制品中结核分枝杆菌检测方法荧光定量PCR法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2016-03-09
Implemented on:	2016-10-01
Superseding:	SN/T 2101-2008 Determination of Mycobacterium tuberculosis for milk and milk products—Real-time PCR method
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	mycobacterium tuberculosis dairy products human mycobacterium tuberculosis real-time pcr method introduction analysis real-time fluorescence pcr instrument
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Introduction

Analysis of the core content of the standard

This standard specifies the standardized process for detecting Mycobacterium tuberculosis in dairy products using fluorescence quantitative PCR instrument, focusing on the specific detection of bovine and human Mycobacterium tuberculosis.

Comparison of technological evolution

Technical indicators	2008 edition	2016 edition	Improvement significance
Sample pretreatment	Single NaOH liquefaction	Triton X-100+ gradient centrifugation	Increase bacterial recovery rate by 30%
Detection sensitivity	10³ CFU/mL	10² CFU/mL	Detection limit reduced by 1 order of magnitude
Quality control system	Double parallel samples	Double parallel samples + confirmation experiment	The false positive rate is reduced to 0.5%

Key operation points

1. Sample preparation

Fresh milk samples need to be centrifuged at 4000r/min, treated with 0.45% NaOH solution for 30 minutes, and digested with proteinase K to effectively break the milk fat barrier.

2. DNA extraction optimization

A new phenol-chloroform extraction step was added, combined with ethanol precipitation, to increase the DNA yield to 85±5%, meeting the detection requirements of real-time fluorescence PCR instrument.

3. Reaction system configuration

A specific probe 5'-FAM-CCGAAGCGGCGCTGGACGAG-TAMRA-3' was used, the amplified fragment was 103bp, and the annealing temperature was strictly controlled at 60℃±0.5℃.

Implementation recommendations

Laboratories need to be equipped with Class II biosafety cabinets and freezing equipment below -20℃
Each batch of tests must include a negative control (sterile water) and a positive control (BCG standard strain)
Critical value samples (28≤Ct≤30) should be tested repeatedly and the template volume should be increased to 2μL
Positive results must be confirmed by strain identification according to GB/T 18645

Application value of the standard

The implementation of this standard shortens the detection cycle of tuberculosis in dairy products from 4-6 weeks of the traditional culture method to within 8 hours, providing efficient technical support for the safety supervision of imported and exported dairy products. The Triton X-100 treatment solution newly added in the 2016 version is particularly suitable for the pre-treatment of high-fat milk samples.

More samples: NB/T 47013.2-2015 YD/T 3934-2021 DL/T 5154-2012

Sample only — not a preview of SN/T 2101-2016

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