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GB/T 27528-2011 English PDF

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GB/T 27528-2011: Real-time RT-PCR for detection of foot and mouth disease virus
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Basic data

Standard ID GB/T 27528-2011 (GB/T27528-2011)
Description (Translated English) Real-time RT-PCR for detection of foot and mouth disease virus
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B41
Classification of International Standard 11.220
Word Count Estimation 9,961
Date of Issue 2011-11-21
Date of Implementation 2012-03-01
Quoted Standard GB/T 6682; GB/T 18088; GB/T 18935; GB 19489; SN/T 1193
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 FMD virus real-time fluorescent RT-PCR detection methods. This standard applies to the detection and identification of FMD virus.

GB/T 27528-2011: Real-time RT-PCR for detection of foot and mouth disease virus

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Real-time RT-PCR for detection of foot and mouth disease virus ICS 11.220 B41 National Standards of People's Republic of China FMD virus real-time fluorescent RT-PCR detection method 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 Annex A, Annex B normative appendix. 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 SZCIQ, Chinese Academy of Inspection and Quarantine, Beijing surplus Jiusi Science and Technology Development Company Limited. The main drafters of this standard. Qinzhi Feng, Lin Xiangmei, Wu Shaojiang, flower Qun Yi, Lu Kang body, Liu, Chen Shukun, Lv Jianjiang, had less spirit, Zhanai Jun, Han Xueqing, Sun Jie, Caochen Fu, Jia Guang Yue, Gao Bo, Chen Bing, Ruanzhou Xi, Mei Lin, Zhang Caihong, TAO. FMD virus real-time fluorescent RT-PCR detection method

1 Scope

This standard specifies the FMD virus in real-time fluorescent RT-PCR detection method. This standard applies to the detection and identification of FMD virus.

2 Normative references

The following documents contain provisions which, through reference in this standard and become the standard terms. For dated references, subsequent Amendments (not including errata content) or revisions do not apply to this standard, however, encourage the parties to the agreement are based on research Whether the latest versions of these documents. For undated reference documents, the latest versions apply to this standard. Laboratory use specifications and test methods GB/T 6682 Analysis GB/T 18088 Entry and Exit Animal Quarantine sampling GB/T 18935 FMD diagnostic techniques GB 19489 General requirements for laboratory biosafety SN/T 1193 genetic testing laboratory technical requirements

3 Abbreviations

The following abbreviations apply to this standard. Ct values. cycle threshold (cyclethreshold).

4 Reagents and equipment supplies

4.1 Reagent Consumables 4.1.1 Unless otherwise indicated, the reagents used in the assay are analytically pure laboratory water should be consistent with GB/T 6682 requirements. 4.1.2 FMDV real-time fluorescent RT-PCR reagents. -20 ℃ to save, see Appendix A. 4.1.3 primer and probe sequences Upstream primer (FMDV-F) sequence. 5'ACTGGGTTTTACAAACCTGTGA3 '. Downstream primer (FMDV-R) sequence. 5'GCGAGTCCTGCCACGGA3 '. TaqMan probes (FMDV-Pb) sequence. 5'TCCTTTGCACGCCGTGGGAC3 ', 5' and 3 'ends are labeled FAM and BHQ. 4.1.4 DEPC-treated water. See Appendix B. Recommended for later commercialization of DEPC-treated DNA and RNA enzyme free water. 4.1.5 chloroform. AR, stored at room temperature. 4.1.6 isopropanol. AR, prior to use pre-cooled to -20 ℃. 4.1.7 75% ethanol. water and ethanol in DEPC formulated before use precooled to -20 ℃. 4.1.8 quartz sand. a commercially available chemically pure, after 121 ℃, 15min autoclave spare. 4.1.9 0.01mol/LPBS, pH7.6 ~ 7.8. See Appendix B. 4.1.10 positive control tissue culture inactivated vaccine virus or inactivated virus, -20 ℃ stored for use. Negative control samples may be healthy pigs Or bovine tissue material. 4.2 The main equipment 4.2.1 PCR fluorescence detector. 4.2.2 high-speed desktop refrigerated centrifuge. temperature controlled to 4 ℃, centrifuged at speeds of up to 12000g above.

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