GB/T 40458-2021Specification of nucleic acid extraction used for high-throughput detection of pathogenic microorganisms (English PDF)

Foreword

This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for Standardization Work Part 1: Structure and Drafting Rules of Standardization Documents" Drafting.

1 Scope

This document specifies the nucleic acid extraction requirements for high-throughput detection of pathogenic microorganisms using second-generation sequencing technology, and describes the corresponding method of extraction.

This document is applicable to the nucleic acid extraction methods of phytopathogenic bacteria, fungi, viruses, and vector-borne bacteria of plants and their products.

2 Normative references

The contents of the following documents constitute the indispensable clauses of this document through normative references in the text. Among them, dated quotations Only the version corresponding to that date is applicable to this document; for undated reference documents, the latest version (including all amendments) is applicable to this document.

GB 19489 Laboratory Biosafety General Requirements

SN/T 2752.4 Self-protection norms for health and quarantine personnel Part 4: Laboratory personnel

SN/T 2776 Management requirements for medical vector monitoring laboratories at frontier ports

SN/T 4278-2015 Operational Regulations for DNA Barcode Identification of Medical Vector Insects at Frontier Ports

SN/T 4625-2016 DNA barcode screening and quality requirements

3 Terms and definitions

There are no terms and definitions that need to be defined in this document.

4 Abbreviations

The following abbreviations apply to this document.

DNA. Deoxyribonucleic Acid (Deoxyribonucleic Acid)

OD. Optical Density (OpticalDensity)

PCR. Polymerase Chain Reaction (PolymeraseChain Reaction)

RNA. Ribonucleic Acid (Ribonucleic Acid)

5 General requirements

Nucleic acid extraction mainly includes two major steps. lysis and purification of biological samples. Lysis is to free the nucleic acid in the sample in the lysis system In the process, purification is to completely separate nucleic acid from other components in the lysis system, such as proteins, polysaccharides, lipids and other tissues or cell components. The process of leaving.

Nucleic acid extraction should ensure the integrity of the primary structure of nucleic acid. Exclude other molecules, such as proteins, polysaccharides, lipids, organic solvents, etc. To ensure the smooth progress of downstream high-throughput detection tests.

During the test, the external environment (such as dust, aerosol, RNase, etc.) should be prevented from contaminating the nucleic acid extraction, as well as the pathogen itself and the test.