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Animal influenza detection -- Method of pyrosequencing for HA and NA in influenza Virus A (H1N1)
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GB/T 27538-2011
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Basic data | Standard ID | GB/T 27538-2011 (GB/T27538-2011) | | Description (Translated English) | Animal influenza detection -- Method of pyrosequencing for HA and NA in influenza Virus A (H1N1) | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | B41 | | Classification of International Standard | 11.220 | | Word Count Estimation | 13,111 | | Date of Issue | 2011-11-21 | | Date of Implementation | 2012-03-01 | | Quoted Standard | GB/T 6682; GB/T 18088; GB 19489; NY/T 541; 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 pyrosequencing technology for the A H1N1 influenza virus and its variants in Mexico HA and NA nucleic acid sequence detection methods. This standard applies to generic type A H1N1 influenza virus type A (H1N1) virus and mutants Mexico nucleic acid detection. |
GB/T 27538-2011: Animal influenza detection -- Method of pyrosequencing for HA and NA in influenza Virus A (H1N1) ---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 pyrosequencing for HA and NA in influenza Virus A (H1N1)
ICS 11.220
B41
National Standards of People's Republic of China
Detecting animal influenza type A H1N1 influenza virus
Pyrosequencing detection HA, NA's
influenzavirusA (H1N1)
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 Shandong Exit Inspection and Quarantine, Chinese Academy of Inspection and Quarantine.
The main drafters of this standard. Xu Biao, Liang beads, Ling Zong Shuai, Zhang Taixiang, Yue Zhiqin, Gao Hongwei, Sun Tao, Fangshao Qing, Lin Xiangmei, Han Xueqing,
Zhu China, Zhang Hexiao single tiger.
Detecting animal influenza type A H1N1 influenza virus
Pyrosequencing detection HA, NA's
1 Scope
This standard specifies the pyrosequencing technique for Type A H1N1 influenza virus mutants in Mexico and the HA and NA nucleic acid
Sequence Detection.
This standard applies to nucleic acid detection type A H1N1 influenza virus type A H1N1 influenza and common virus variants Mexico.
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/T 18088 Entry and Exit Animal Quarantine sampling
GB 19489 General requirements for laboratory biosafety
NY/T 541 animal disease laboratory sampling method
SN/T 1193 genetic testing laboratory technical requirements
3 Abbreviations
The following abbreviations apply to this document.
ATP. adenosine triphosphate (adenosinetriphosphate)
DNA. deoxyribonucleic acid (deoxyribonucleicacid)
DEPC. diethyl pyrocarbonate (diethylpyrocarbonate)
PBS. phosphate buffered saline (phosphatebelancedsolution)
Pyrosequencing. Pyrosequencing
RT-PCR. reverse transcription-polymerase chain reaction (reversetranscriptionpolymerasechainreaction)
RNA. ribonucleic acid (ribonucleicacid)
Taq enzyme. TaqDNA polymerase
Principle 4
Pyrosequencing is a single-stranded DNA template was amplified by PCR and sequencing primer hybridization, and DNA polymerase, ATP sulfuric acid enzymes, fluorescent
Luciferase, adenosine triphosphate and dual phosphatase substrate (APS), fluorescein incubation, each added four kinds of dNTPs (dATP, dTTP, dCTP,
of dGTP), and as the template pair, this end of the primer dNTP form a covalent bond, a group releasing pyrophosphate (PPi); ATP sulfurylase in
The case of the presence of a catalytic APS pyrophosphate formed ATP, ATP driven luciferase-mediated oxidation of luciferin-converting phosphor to oxide phosphor
Optical element emits visible light signal proportional to the amount of ATP; the optical signal from the CCD detector (CCD) and collected by the software into peaks.
Nucleotide number is proportional to the peak height of the response signal in each optical incorporated. ATP and unincorporated dNTP by the adenosine triphosphate diphosphate
Enzymatic degradation, quenched optical signal regeneration and reaction system.
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