T/YNANT 08-2024 Active Group standards

T/YNANT 08-2024 The Multi-target Real-Time Quantitative RT-qPCR Detection Method for Avian Influenza Virus H5, H7, and H9 Subtypes

Publish Date: 2024-02-06 Implement Date: 2024-02-16 For services related to genuine standard inquiry, procurement, translation, and other related services in China, please Contact Us

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Basic Information

Standard Code: T/YNANT 08-2024

Standard Type: Group standards

Standard Status: Active

is_force_gb: no

CCS Name: -

ICS Name: Veterinary medicine

Publish Date: 2024-02-06

Implement Date: 2024-02-16

Scope

Scope: This document specifies the reagents and materials, instruments and equipment, pre-experiment preparations, operating procedures, and result determination for the real-time fluorescence quantitative RT-qPCR detection of the H5, H7, and H9 subtypes of the avian influenza virus (AIV) in poultry and wild birds, as well as in chicken embryo allantoic fluid, excreta, secretions, and environmental samples. Main technical content: Based on the specific sequences of the shared genes of the avian influenza virus, a pair of specific primers and a specific fluorescent dual-labeled probe are synthesized. The 5' and 3' ends of the probe are labeled with different fluorescent dyes, such as the 5' end labeled with FAM fluorescent dye, which emits fluorescence that can be detected by the detection instrument and is called the reporter fluorescent group (represented by R); the 3' end is generally labeled with TAMRA fluorescent dye, which can absorb the fluorescence signal emitted by the reporter fluorescent group at the 5' end at close range and is called the quenching fluorescent group (represented by Q). When the PCR reaction is in the annealing stage, a pair of primers and a probe simultaneously bind to the target gene fragment. At this time, the fluorescence signal emitted by the R group on the probe is absorbed by the Q group, and the instrument cannot detect the fluorescence signal emitted by R; when the PCR reaction proceeds to the extension stage, Taq enzyme, guided by the primers, uses four nucleotides as substrates to synthesize a new chain along the template strand according to the principle of base pairing; when the chain extension reaches the probe binding site, it is hindered by the probe and cannot continue. At this time, Taq enzyme hydrolyzes the probe into a single nucleotide, eliminates the obstacle, and at the same time, the R group labeled on the probe is released and detected by the detection instrument; under the action of Taq enzyme, the extension proc