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GB/T 27540-2011: Method of the real-time RT-PCR for the detection of classical swine fever virus
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivered in | Standard Title (Description) | Status | PDF |
| GB/T 27540-2011 | English | 209 |
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Method of the real-time RT-PCR for the detection of classical swine fever virus
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GB/T 27540-2011
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Basic data | Standard ID | GB/T 27540-2011 (GB/T27540-2011) | | Description (Translated English) | Method of the real-time RT-PCR for the detection of classical swine fever virus | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | B41 | | Classification of International Standard | 11.220 | | Word Count Estimation | 9,969 | | Date of Issue | 2011-11-21 | | Date of Implementation | 2012-03-01 | | 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 classical swine fever virus (Classical swine fever virus, CSFV) real-time fluorescent RT-PCR detection methods. This standard applies to classical swine fever in the diagnosis and monitoring of live pigs for their organs, blood, excrement, and cell culture CSFV nucleic acid detection. | GB/T 27540-2011: Method of the real-time RT-PCR for the detection of classical swine fever virus---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.
Method of the real-time RT-PCR for the detection of classical swine fever virus
ICS 11.220
B41
National Standards of People's Republic of China
Classical swine fever 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 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. Chinese Veterinary Drug Control.
The main drafters of this standard. Wang Qin, Wangtai Jian, Xu Lu, Fanxue Zheng, Liu, Jiang Chunyan, Zhao Qizu, Ning Yi Bao, Zouxing Qi.
Classical swine fever virus real-time fluorescent RT-PCR detection method
1 Scope
This standard specifies the swine fever virus (Classicalswinefevervirus, CSFV) real-time fluorescent RT-PCR detection method.
This standard applies to the diagnosis and monitoring of classical swine fever for live pigs and their organs, blood, excrement, and cell culture CSFV nuclear
Acid test.
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.
People's Republic of China Ministry of Agriculture Bulletin No. 302 veterinary laboratory biosafety management practices
3 Abbreviations
The following abbreviations apply to this document.
CSFV. classical swine fever virus (Classicalswinefevervirus)
Ct values. reach the threshold number of cycles (cyclethreshold)
DEPC. diethyl pyrocarbonate (diethylpyrocarbonate)
HSTaq enzyme. TaqDNA hot start polymerase (HSTaqDNApolymerase)
RNA. ribonucleic acid (ribonucleicacid)
Fluorescent RT-PCR. reverse transcriptase polymerase chain reaction fluorescence (realtimefluorescentquantitativereverse
transcriptionpolymerasechainreaction)
4 Reagents and materials
Unless otherwise indicated, the reagents used were of analytical grade; all reagents were used without RNA enzyme dispensing container.
4.1 Trizol. RNA extraction reagent, appearance of pink to brown bottle packing, stored at 4 ℃ ~ 8 ℃.
4.2 chloroform. 4 ℃ pre-cooling.
4.3 Isopropanol. 4 ℃ pre-cooling.
4.4 75% ethanol. anhydrous ethanol with a new open and DEPC water preparation, -20 ℃ pre-cooling.
4.5 DEPC water. deionized water 0.1PC, 37 ℃ role 1h, (121 ± 2) ℃, autoclave 15min.
4.6 RNA inhibitor (30U/μL).
4.7 10 × PCRbuffer (10mmol/LpH8.3Tris-HCl, 50mmol/LKCl, 1.5mmol/LMgCl2).
4.8 dNTP (2.5mmol/L).
4.9 Primers for RT-PCR reactions the concentration is 10μmol/L, the concentration of the probe 5μmol/L, which sequence is as follows.
The upstream primer F. 5-TACAGGACAGTCGTCAGTAGTTCGA-3
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