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GB/T 27539-2011 English PDF

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GB/T 27539-2011: Animal influenza detection -- Method of real-time RT-PCR for detection of influenza virus A
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GB/T 27539-2011English229 Add to Cart 3 days [Need to translate] Animal influenza detection -- Method of real-time RT-PCR for detection of influenza virus A Valid GB/T 27539-2011

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Basic data

Standard ID GB/T 27539-2011 (GB/T27539-2011)
Description (Translated English) Animal influenza detection -- Method of real-time RT-PCR for detection of influenza virus A
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B41
Classification of International Standard 11.220
Word Count Estimation 10,188
Date of Issue 2011-11-21
Date of Implementation 2012-03-01
Quoted Standard GB/T 6682; GB 19489; GB/T 19495.2; GB/T 27401; SN/T 1330
Regulation (derived from) Announcement of Newly Approved National Standards No. 18 of 2011
Issuing agency(ies) General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Standardization Administration of the People's Republic of China
Summary This standard specifies the type A influenza virus Universal fluorescent RT-PCR detection method of operation. This standard applies to live animals and their products in the detection of influenza A virus.

GB/T 27539-2011: Animal influenza detection -- Method of real-time RT-PCR for detection of influenza virus A


---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Animal influenza detection. Method of real-time RT-PCR for detection of influenza virus A ICS 11.220 B41 National Standards of People's Republic of China Detecting animal influenza type A influenza virus General fluorescent RT-PCR detection method influenzavirusA Issued on. 2011-11-21 2012-03-01 implementation Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China Standardization Administration of China released

Foreword

This standard was drafted in accordance with GB/T 1.1-2009 given rules. The standard proposed by the People's Republic of China Ministry of Agriculture. This standard by the National Standardization Technical Committee Animal Epidemic Prevention (SAC/TC181) centralized. This standard was drafted. People's Republic of China Beijing Entry-Exit Inspection and Quarantine Bureau, People's Republic of China Guangdong Entry-Exit Inspection and Quarantine Bureau, the Shenzhen-based horses Biological Engineering Co., Ltd. The main drafters of this standard. Wang Lin, Lin Zhixiong, Zhou Qi, Gao Zhiqiang, Chen Ru, Qiao Caixia, Pu Jing, Yang, Liang beads. Detecting animal influenza type A influenza virus General fluorescent RT-PCR detection method

1 Scope

This standard specifies the method of operation of influenza A virus common fluorescent RT-PCR detection. This standard applies to live animals and their products in the detection of influenza A virus.

2 Normative references

The following documents for the application of this document is essential. For dated references, only the dated version suitable for use herein Member. For undated references, the latest edition (including any amendments) applies to this document. Laboratory use specifications and test methods GB/T 6682 Analysis GB 19489 General requirements for laboratory biosafety GB/T 19495.2 technical requirements for GMO testing laboratories GB/T 27401 laboratory quality control standards of Animal Quarantine SN/T 1330 import and export of raw and cooked fur inspection procedures

3 Abbreviations

The following abbreviations apply to this document. Cycles (Ctvalue) fluorescence signal within each reaction tube reaches a set threshold when experienced. Ct values DEPC. diethyl pyrocarbonate (diethylpyrocarbonate) PBS. phosphate buffer (recipe see Appendix A) (phosphatebuffersaline) RNA. ribonucleic acid (ribonucleicacid) RT-PCR. reverse transcription - polymerase chain reaction (reversetranscriptionpolymerasechainreaction) Principle 4 Influenza virus (influenzavirus) genus Orthomyxoviridae, depending on the antigenic characteristics and genetic characterization of the influenza A virus into (A), B (B), propyl (C) type. B variability weak, hepatitis antigen relatively stable, only infects humans; A (A) antigen variability of the strongest, Infect humans and other animals, often causing a worldwide pandemic. Encoding matrix proteins M gene of influenza A virus, and there are more conservative. Using Taqman technology, in particular for the M gene sequence, synthesized a pair of specific primers and a specific dual-labeled fluorescent probes, probe 5-pin 'end labeled with a reporter fluorophore (R), 3' end labeled fluorophore quencher (Q). In the PCR annealing stage, a pair of primers and a probe Simultaneously with the target gene fragment binds, the fluorescence signal emitted by the probe R group is absorbed by Q group, the instrument can not detect the emitted R Fluorescence signal; in PCR extension phase, Taq polymerase under the guidance of the primer along the template strand synthesis of new chain, the chain will be extended when the probe binds to When parts hindered the probe can not continue, Taq enzyme plays 5 '→ 3' exonuclease function as a single nucleotide hydrolysis probes labeled Written in the R groups on the probe freed, Q absorbed by the detector being fluorescence received by R is not issued, as the PCR reaction Performed, PCR product and the amount of fluorescence signal exhibits a proportional relationship.

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