GB/T 36848-2018 in English
VALIDDetection and identification of Avocado sunblotch viroid
- Issued on:2018-09-17
- Implemented on:2019-04-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$156.00
《GB/T 36848-2018鳄梨日斑类病毒检疫鉴定方法》由TC271（全国植物检疫标准化技术委员会）归口，主管部门为国家标准化管理委员会。
Introduction
Interpretation of the Standard Method for Quarantine Identification of Avocado Day Spot Viroid
| Dimensions | Content |
|---|---|
| Scope of the Standard | Applicable to the quarantine identification of avocado seedlings, seeds and fruits, using RT-PCR and R-PAGE methods. |
| Detection Principle | Identification is based on RNA molecular weight differences and sequence characteristics to ensure accuracy. |
| Instruments | PCR instrument, electrophoresis instrument, gel imaging system, etc., to ensure experimental accuracy and repeatability. |
Background of Standard Formulation and Analysis of Technological Evolution
Background Introduction:
Avocado day spot virus-like (ASBVd) is a pathogen that seriously harms avocado cultivation. It has various transmission routes, including seeds, grafting and mechanical transmission. The formulation of this standard aims to provide unified technical specifications for plant quarantine departments to ensure accurate identification and control of virus spread.
Technological Evolution:
From traditional pathological methods to the application of molecular biology techniques, detection methods have undergone significant improvements. The introduction of RT-PCR and R-PAGE methods has greatly improved the sensitivity and specificity of detection, providing strong support for quarantine work.
Professional interpretation of avocado day spot viroid detection method
Detailed explanation of RT-PCR detection method
RT-PCR is one of the core technologies of this standard. The target fragment of 247bp is amplified by a specific primer pair (ASBVdf and ASBVdr), and the specificity of the product is verified by agarose gel electrophoresis.
- Primer design: ASBVdf (5'-AGTTCACTCGTCTTCAATCTC-3') and ASBVdr (5'-CTGAAGAGACGAAGTGATCAA-3'), the amplified product length is 247bp.
- Operation process: Includes RNA extraction, reverse transcription and PCR amplification, and the results are finally confirmed by electrophoresis.
Reciprocating two-dimensional polyacrylamide gel electrophoresis (R-PAGE)
R-PAGE is a high-resolution RNA analysis technique that separates RNA molecules by two-dimensional electrophoresis, using bromophenol blue and xylene cyanol as tracers.
- Gel preparation:6% polyacrylamide gel with 10×TBE buffer.
- Electrophoresis steps:The first dimension electrophoresis separates RNA molecules, and the second dimension electrophoresis further confirms the differences in molecular weight and mobility.
Implementation recommendations
Laboratory capacity building
It is recommended that all plant quarantine agencies be equipped with advanced molecular biology equipment, including PCR instruments, electrophoresis systems and gel imagers, and conduct regular technical training.
Sample management optimization
Establish a standardized sample collection and preservation process to ensure the integrity of samples during transportation and storage. It is recommended to use low temperature freezing (-80°C) to preserve positive samples.
Conclusion
GB/T 36848—2018 standard provides a scientific basis and technical specifications for the quarantine and identification of avocado day spot viroid. The RT-PCR and R-PAGE methods, combined with a strict sample management process, can effectively control the risk of disease transmission.
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