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SN/T 3327-2012: Quarantine protocal for classic swine fever virus by loop-mediated isothermal amplification
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| SN/T 3327-2012 | English | 209 |
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Quarantine protocal for classic swine fever virus by loop-mediated isothermal amplification
| Valid |
SN/T 3327-2012
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Standard similar to SN/T 3327-2012 SN/T 5268 Basic data | Standard ID | SN/T 3327-2012 (SN/T3327-2012) | | Description (Translated English) | Quarantine protocal for classic swine fever virus by loop-mediated isothermal amplification | | Sector / Industry | Commodity Inspection Standard (Recommended) | | Word Count Estimation | 8,839 | | Quoted Standard | GB/T 18088; GB 19489-2008 | | Regulation (derived from) | National Quality Inspection (2012) 777; industry standard filing Notice 2013 No. 4 (No. 160 overall) | | Issuing agency(ies) | General Administration of Customs | | Summary | This standard specifies the method of operation of classical swine fever virus reverse transcriptase loop-mediated isothermal nucleic acid amplification testing. This standard applies to rapid screening of classical swine fever virus. | SN/T 3327-2012: Quarantine protocal for classic swine fever virus by loop-mediated isothermal amplification
---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.
Quarantine protocal for classic swine fever virus by loop-mediated isothermal amplification
People's Republic of China Entry-Exit Inspection and Quarantine Standards
Classical swine fever virus by reverse transcription loop-mediated isothermal
Nucleic acid amplification tests
Quarantineprotocalforclassicswinefevervirusbyloop-I mediated
Issued on. 2012-12-12
2013-07-01 implementation
People's Republic of China
The State Administration of Quality Supervision, Inspection and Quarantine released
Foreword
This standard was drafted in accordance with GB/T 1.1-2009 given rules.
This standard is proposed and managed by the National Certification and Accreditation Administration Committee.
This standard was drafted. People's Republic of China Shanghai Entry-Exit Inspection and Quarantine Bureau.
The main drafters of this standard. Li Jian, Xiong Wei, Zhang Qiang, Jiang Jing, Qiu Lu, Wang Qiao whole, Huang Zhongrong, Hu Yongqiang.
Classical swine fever virus by reverse transcription loop-mediated isothermal
Nucleic acid amplification tests
1 Scope
This standard specifies the method of operation of classical swine fever virus by reverse transcription loop-mediated isothermal nucleic acid amplification detection.
This standard applies to rapid screening of classical swine fever 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.
GB/T 18088 Entry and Exit Animal Quarantine sampling
GB 19489-2008 General requirements for laboratory biosafety
3 Overview
Classical swine fever (classicalswinefever, CSF), is a kind of emergency by the classical swine fever virus (classicalswinefevervirus, CSFV) caused
Resistance, heat resistance, highly contagious disease of swine breeding industry very serious harm. CSFV genotype positive strand RNA, genome about
123kb, contains only one large open reading frame (ORF). The standard use of loop-mediated isothermal nucleic acid amplification at a constant temperature
Fast, efficient and specifically amplify located fever virus 5 'untranslated region (5'-UTR) of the target sequence, so as to achieve rapid detection purposes.
4 Reagents and materials
4.1 BstDNA polymerase (large fragment). enzyme concentration was 8U/μL, -20 ℃, avoid repeated freezing and thawing.
4.2 10 times ThermoPol polymerase buffer concentrate. -20 ℃ to save.
4.3 magnesium sulfate (MgSO4). concentration of 25mmol/L, -20 ℃ preservation.
4.4 AMV reverse transcriptase. reverse transcriptase concentration of 5U/μL, -20 ℃, avoid repeated freezing and thawing.
4.5 betaine solution (Betaine). concentration 5mol/L, -20 ℃, avoid repeated freezing and thawing.
4.6 dNTPs. containing dATP, dGTP, dCTP, dTTP each 10mmol/L, -20 ℃ preservation.
4.7 LAMP primer. primer dubbed the concentration of 25μmol/L, -20 ℃ stored for use. Its sequence is as follows.
--- Being outwardly primer (CSFV-F3). 5'-AGC-TCC-CTG-GGT-GGT-CTA-3 ';
--- Reverse outer primer (CSFV-B3). 5'-CCT-AAT-AGT-GGG-CCT-CTG-CA-3 ';
--- Within the forward primer (CSFV-FIP). 5'-GCC-CTC-GTC-CAC-ATA-GCA-TCT-CAG-TAC-AGG-ACA-
GTC-GTC-AGT-3 ';
--- Within the reverse primer (CSFV-BIP). 5'-GCC-CAA-GAC-ACA-CCT-TAA-CCC-TTC-AGG-TCG-TAC-
TCC-CAT-CAC-3 ';
--- Forward loop primer (CSFV-LF). 5'-GGG-CTT-CTG-CTC-ACG-TCG-3 ';
--- Reverse loop primer (CSFV-LB). 5'-GGT-CGC-TAG-GGT-GAA-ATC-3 '.
4.8 SYBRGreenⅠ fluorescent dye solution. -20 ℃ to save.
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