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SN/T 4799-2017 English PDF

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SN/T 4799-2017: Detection of tuberculosis pathogenic organisms of animals. DHPLC method
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Basic data

Standard ID SN/T 4799-2017 (SN/T4799-2017)
Description (Translated English) Detection of tuberculosis pathogenic organisms of animals. DHPLC method
Sector / Industry Commodity Inspection Standard (Recommended)
Classification of Chinese Standard B41
Word Count Estimation 10,115
Date of Issue 2017-07-21
Date of Implementation 2018-03-01
Quoted Standard GB 19489
Regulation (derived from) State-Quality-Inspection-Accreditation (2017) 337
Issuing agency(ies) General Administration of Customs
Summary This standard specifies the technical requirements and operational specifications for the detection of Mycobacterium tuberculosis complex, Mycobacterium tuberculosis, and Mycobacterium bovis multiplex PCR combined with denaturing high performance liquid chromatography (mPCR-DHPLC). This standard is applicable to the rapid detection of Mycobacterium tuberculosis complex, Mycobacterium tuberculosis and Mycobacterium bovis in clinical samples such as bacterial culture, animal tissue, blood sample and sputum.

SN/T 4799-2017: Detection of tuberculosis pathogenic organisms of animals. DHPLC method

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Detection of tuberculosis pathogenic organisms of animals.DHPLC method People's Republic of China entry and exit inspection and quarantine industry standards Method for detecting pathogens of animal tuberculosis Denaturing high performance liquid chromatography DHPLCmethod Released on.2017-07-21 2018-03-01 implementation People's Republic The General Administration of Quality Supervision, Inspection and Quarantine issued Entry-Exit Inspection and Quarantine Industry Standard Method for detecting pathogens of animal tuberculosis Denaturing high performance liquid chromatography China Standard Press Publishing First edition, June.2018

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard is proposed and managed by the National Certification and Accreditation Administration. This standard was drafted. Guangdong Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China. The main drafters of this standard. Liu Zhiling, Chen Ru, Wang Ying, Wu Xiaowei, Zhu Daozhong, Duan Yanyu, Lin Zhixiong. Method for detecting pathogens of animal tuberculosis Denaturing high performance liquid chromatography

1 Scope

This standard specifies the multiplex PCR combined with denaturing high performance liquid chromatography of Mycobacterium tuberculosis complex, Mycobacterium tuberculosis and Mycobacterium bovis (mPCR-DHPLC) Technical requirements and operating specifications for the detection method. This standard is applicable to the rapid detection of Mycobacterium tuberculosis complexes and tuberculosis in clinical samples such as bacterial cultures, animal tissues, blood samples and sputum. Mycobacteria, Mycobacterium bovis.

2 Normative references

The following documents are indispensable for the application of this document. For dated references, only the dated version applies to this article. Pieces. For undated references, the latest edition (including all amendments) applies to this document. GB 19489 General requirements for laboratory biosafety

3 Abbreviations

The following abbreviations apply to this document. DHPLC. denaturing high-performance liquid chromatography (denaturing high-performance liquid chromatography). dNTP. deoxyribonuclesosdetriphosphate. PBS. phosphate buffer solution. mPCR. multiplex-polymerase chain reaction. TEAA. Triethylammonium acetate.

4 Overview

Human and animal tuberculosis involves multiple pathogens or pathogen complexes. Mycobacterium tuberculosis complex (Mycobacteriumtuber- Culosiscomplex (MTC) is a tuberculosis pathogen that infects humans and mammals, including Mycobacterium tuberculosis and Mycobacterium bovis, Africa Mycobacterium and M. variabilis. Among them, M. tuberculosis and M. bovis are mainly caused by infection of humans and livestock. With MTC Corresponding to a wide variety of non-tuberculous mycobacteria (NTM). Clinical symptoms and pathological changes in patients with NTM infection and MTC The resulting tuberculosis is very similar, but most NTMs are naturally resistant to tuberculosis drugs. In addition, NTM infection can cause tuberculin False positives in intradermal allergy or serological tests. Simultaneous differentiation of knots by multiplex PCR combined with denaturing high performance liquid chromatography Mycobacterium tuberculosis complex and non-tuberculous mycobacteria review group, and identification of Mycobacterium tuberculosis and Mycobacterium bovis The principle and operation of the mPCR reaction are the same as those of ordinary PCR. Add multiple pairs of specific primers in the same PCR reaction system, Multiple DNA fragments are amplified from multiple DNA templates or different regions of the same template. DHPLC analysis technique Technique, in a high-pressure closed liquid phase flow path, a DNA sample is automatically injected and carried under a buffer to flow through a DNA separation column, a DNA fragment The negatively charged phosphate group in the molecule interacts with the positively charged amino group in the TEAA molecule to attract each other while TEAA The three ethyl groups in the molecule interact with each other by the hydrophobic interaction of the alkyl group on the surface of the stationary phase C18, through the gradient of acetonitrile in the mobile phase. Elution separates DNA fragments of different sizes. The isolated DNA sample is detected by ultraviolet or fluorescence. According to the length of the DNA amplified fragment

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