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GB/T 31805-2015: Detection and identification of Cowpea severe mosaic virus
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| GB/T 31805-2015 | English | 339 |
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Detection and identification of Cowpea severe mosaic virus
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GB/T 31805-2015
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Basic data | Standard ID | GB/T 31805-2015 (GB/T31805-2015) | | Description (Translated English) | Detection and identification of Cowpea severe mosaic virus | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | B16 | | Classification of International Standard | 65.020.01 | | Word Count Estimation | 16,178 | | Date of Issue | 2015-07-03 | | Date of Implementation | 2015-11-27 | | Regulation (derived from) | National Standard Announcement 2015 No.22 | | 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 cowpea severe mosaic virus quarantine identified serology, molecular biology and detection methods. This Standard applies may carry heavy cowpea mosaic virus in a plant propagation material (seeds, plants) and for quarantine and identification products. | GB/T 31805-2015: Detection and identification of Cowpea severe mosaic 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.
Detection and identification of Cowpea severe mosaic virus
ICS 65.020.01
B16
National Standards of People's Republic of China
Cowpea severe mosaic virus quarantine and identification methods
Issued on. 2015-07-03
2015-11-27 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.
Please note that some of the content of this document may involve patents. Distribution of this document
Institutions do not assume the responsibility to identify these patents.
This standard by the National Standardization Technical Committee of Plant Quarantine (SAC/TC271) and focal points.
This standard was drafted. Xiamen, People's Republic of China Exit Inspection and Quarantine, People's Republic of China Jiangsu Entry-Exit Inspection and
Quarantine Bureau.
The main drafters of this standard. Chen Qing, Li Bin, Fang Zhipeng, Huang Feng, Liao Furong, Chen Yun, Lin Shiming.
Cowpea severe mosaic virus quarantine and identification methods
1 Scope
This standard specifies the cowpea severe mosaic virus quarantine identified serology, molecular biology and detection methods.
This standard applies may carry heavy cowpea mosaic virus in a plant propagation material (seeds, plants) and for quarantine and identification products.
2 Equipment and Reagents
2.1 Equipment
PCR, quantitative PCR, autoclaves, ice maker, ultra-low temperature freezer, a conventional refrigerator, whirlpool oscillator, electrophoresis, gel imaging system
EC, micro-mills, microplate reader, washer, microbalance (sense of volume. 0.001g), horizontal electrophoresis tank, desktop high-speed refrigerated centrifuge, water bath,
pH meter, pipettes (1000μL, 200μL, 20μL, 2μL), 96 well microtiter plates, mortar or the like; pest control greenhouse.
2.2 Reagents
Unless otherwise specified, all reagents were of analytical grade. PCR buffer, dNTPs (dATP, dTTP, dCTP, dGTP), DNA
Polymerase, primers and probes, agar, ELISA detection reagent in Appendix B.
Preparation of test sample 3
3.1 Seed
Picked deformity seed sown in sterilized soil, grow to be 3 to 4 leaves, the plants will show symptoms of number of plants not showing symptoms
Group (10 per group) and number. Collecting leaves divided into two parts, respectively, as required for enzyme-linked assays and molecular testing.
3.2 Plant
Symptoms (such as blade malformation, mosaic, mottled, etc.) number of plants individually tested. No symptoms of the grouping and number detection, grouping
And detection methods with 3.1. Double antibody sandwich enzyme-linked immune assay.
4 Detection and Identification
4.1 pairs immunosorbent assay antibody sandwich enzyme-linked
Sample preparation was added to the supernatant which had been coated CPSMV antibody ELISA plate, make the DAS-ELISA. Each sample was added to parallel
Two holes. Let the negative and positive controls [healthy plant tissue as negative control, infected plant tissue CPSMV to make positive
According to which the negative control and the type of material (such as seed or blade) should be consistent with the test sample], sample extraction buffer as control, not
With detection reagents or kits according to the instructions. For details, see Appendix B.
4.2 RT-PCR
Total RNA was extracted from the samples and controls, after reverse transcription synthesis of cDNA, PCR amplification. Healthy plant tissue as negative for
Photos, infection of plant tissue CPSMV as positive control, with ultra-pure water as control. For details, see Appendix C.
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