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(Odontoglossum ringspot virus quarantine and identification methods)
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SN/T 4077-2014
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Standard similar to SN/T 4077-2014 SN/T 5268 Basic data | Standard ID | SN/T 4077-2014 (SN/T4077-2014) | | Description (Translated English) | (Odontoglossum ringspot virus quarantine and identification methods) | | Sector / Industry | Commodity Inspection Standard (Recommended) | | Date of Issue | 12/1/2014 | | Date of Implementation | 5/1/2015 | | Regulation (derived from) | State-Quality-Inspection-accreditation [2014] 614 | | Issuing agency(ies) | General Administration of Customs | | Summary | This standard specifies the molecular biology and immunology methods Odontoglossum ringspot virus quarantine identified. This standard applies to quarantine and identification of orchid seedlings and plant products may toothed ringspot virus. | SN/T 4077-2014: (Odontoglossum ringspot virus quarantine and identification methods)---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.
(Odontoglossum ringspot virus quarantine and identification methods)
People's Republic of China Entry-Exit Inspection and Quarantine Industry Standard
Method for quarantine and identification of tooth ring spot virus
Released on.2014-11-19
2015-05-01 implementation
People's Republic
The General Administration of Quality Supervision, Inspection and Quarantine issued
Foreword
This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
This standard is proposed and managed by the National Certification and Accreditation Administration.
This standard was drafted. Xiamen Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China, Jiangsu Entry-Exit Inspection and Quarantine
Bureau, Xiamen Plant Protection and Quarantine Station.
The main drafters of this standard. Chen Qing, Huang Feng, Li Bin, Liao Furong, Xie Yizhen, Chen Hongyun, Lin Shiming.
Method for quarantine and identification of tooth ring spot virus
1 Scope
This standard specifies the immunological and molecular biological detection methods for quarantine identification of tooth ringworm.
This standard applies to the quarantine and identification of orchid seedlings and plant products that may have tooth ring spot virus.
2 tooth ring spot virus basic information
Scientific name. odontoglossumringspotvirus
Abbreviation. ORSV
Taxonomic status. a member of the Tobamovirus.
ORSV mainly transmits through mechanical transmission, and peach aphid can transmit drugs.
See Appendix A for details.
3 Principle of the method
According to the specific reaction between the tooth ring virus and the antibody, the orchid leaves were tested by ELISA; according to the virus genome
The specificity of PCR was detected, and the results were determined by the size of the electrophoresis strip.
4 instruments, facilities and reagents
4.1 Instruments and facilities
Enzyme-linked detector, electronic balance (inductive quantity 0.0001g), PCR instrument, electrophoresis instrument, electrophoresis tank, gel imaging system, constant temperature water bath, low temperature ice
Box, ordinary refrigerator, centrifuge, etc.; micropipette (2.5μL, 10μL, 20μL, 100μL,.200μL, 1000μL); enzyme plate, mortar
Etc; anti-insect greenhouse.
4.2 Reagents
Enzyme-linked detection reagents (see Appendix B), RT-PCR detection reagents (see Appendix C), IC-RT-PCR detection reagents (see Appendix D).
5 Quarantine and identification
5.1 Sample preparation
The samples were observed and the seedling numbers with symptoms (eg, leaf deformity, mosaic, mottle, etc.) were detected separately. Group without symptoms and numbered
Test, group, 10 strains into one group, the collected leaves are divided into two parts, which are used for enzyme-linked detection and molecular biological detection, respectively, less than 10 strains per plant
Sampling for testing.
5.2 Detection method
5.2.1 Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA)
The prepared sample supernatant was added to a 96-well enzyme-linked plate coated with an ORSV antibody, and subjected to DAS-ELISA. Each sample
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