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GB/T 22915-2008: Protocol of universal fluorogenic RT-PCR for foot and mouth disease virus
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivered in | Standard Title (Description) | Status | PDF |
| GB/T 22915-2008 | English | 279 |
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Protocol of universal fluorogenic RT-PCR for foot and mouth disease virus
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GB/T 22915-2008
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Basic data | Standard ID | GB/T 22915-2008 (GB/T22915-2008) | | Description (Translated English) | Protocol of universal fluorogenic RT-PCR for foot and mouth disease virus | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | B41 | | Classification of International Standard | 11.220 | | Word Count Estimation | 12,150 | | Date of Issue | 2008-12-31 | | Date of Implementation | 2009-05-01 | | Regulation (derived from) | National Standard Approval Announcement 2008 No.25 (Total No.138) | | 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 method of operation of FMD virus fluorescent RT-PCR detection. This standard is applicable to the detection of animals and animal products FMD virus. | GB/T 22915-2008: Protocol of universal fluorogenic RT-PCR for foot and mouth disease 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.
Protocol of universal fluorogenic RT-PCR for foot and mouth disease virus
ICS 11.220
B41
National Standards of People's Republic of China
FMDV fluorescent RT-PCR detection method
Posted 2008-12-31
2009-05-01 implementation
Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China
Standardization Administration of China released
Foreword
The standard reference to the OIE (OIE) "Terrestrial Animals Manual of diagnostic tests and vaccines (mammals, birds and bees)"
(5th edition).
The Standard Appendix A, Appendix C is a normative appendix, Appendix B is an informative annex.
The standard proposed by the People's Republic of China Ministry of Agriculture.
This standard by the National Standardization Technical Committee on Animal Epidemic Prevention.
This standard was drafted. People's Republic of China, Shenzhen Entry-Exit Inspection and Quarantine Yunnan People's Republic of China
Bureau, Chinese Academy of Inspection and Quarantine.
The main drafters of this standard. flower Qun Yi, Yang Yunqing, Qinzhi Feng, Zhou Xiaoli, Lu body Kang Dong Jun, TAO, Ruanzhou Xi, Yi Ye excellent, Lin Xiangmei,
Wu Shaojiang.
FMDV fluorescent RT-PCR detection method
1 Scope
This standard specifies the method of operation FMDV fluorescent RT-PCR detection.
This standard applies to animals and animal products detect FMD virus.
2 Acronyms
The following abbreviations apply to this standard.
2.1 fluorescent RT-PCR
Fluorescence reverse transcription - polymerase chain reaction.
The number of cycles when fluorescence signal of each reaction tube reaches a set threshold experienced.
2.3 RNA
RNA.
2.4 DEPC
Dicarbonate phospholipids.
2.5 PBS
Phosphate buffered saline, recipe see Appendix A.
2.6 T Rao enzyme q
T Rao q DNA polymerase.
2.7 FMD Vp
FMD virus.
Principle 3
According fragments from various types of FMD virus common gene specific sequence, A pair of universal primers and a common
Specific probe. Fluorescent probes 5 'fluorescein labeled FAM, 3' TAMRA labeled fluorescein, the 3 'end of the quencher in close
Can absorb within the 5 'fluorophore reporter fluorescent signal emitted. However, in amplifying, since the T q Rao enzyme 5 '→ 3' exonuclease activity, extension
When the fluorescent probe, which was cut to two groups separated, quenching effect disappears, the fluorescence signal. Therefore, by detecting the fluorescence signal
Template nucleic acid is detected.
4 Materials and Reagents
4.1 Instruments and Equipment
4.1.1 RT-PCR fluorescence detector.
4.1.2 high-speed desktop refrigerated centrifuge (centrifugal speed 12000r/min or more).
4.1.3 tabletop centrifuge (centrifugal speed 3000r/min).
4.1.4 Mixer.
4.1.5 refrigerator (2 ℃ ~ 8 ℃ and -20 ℃ two kinds).
4.1.6 Micro adjustable pipette (5μL, 10μL, 100μL, 1000μL) and supporting filter tips.
4.1.7 Eppendorf tubes (1.5mL), a transparent thin-walled PCR tube (0.2mL).
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