GB/T 36812-2018 in English
VALIDMolecular detection method of Potato yellow dwarf virus
- Issued on:2018-09-17
- Implemented on:2019-04-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$214.00
《GB/T 36812-2018马铃薯黄矮病毒分子生物学检测方法》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
GB/T 36812—2018 Standard Overview
This standard specifies the molecular biological detection method for Potato yellow dwarf virus (PYDV) and is applicable to the quarantine and identification of plants and plant products that may carry the virus. The standard is proposed and managed by the National Technical Committee for Plant Quarantine and Standardization.
Comparison of Standard Frameworks
| Chapter | Content Overview | Keywords |
|---|---|---|
| 1 Scope | Specify the scope of application of the detection method | Potato yellow dwarf virus, Molecular biology detection |
| 5 Instruments, Utensils and Reagents | List the required equipment and reagents | PCR instrument, Fluorescence PCR instrument, Total RNA extraction reagent |
| 7 Detection and Identification | Detailed detection process and judgment criteria | RT-PCR, Real-time fluorescence RT-PCR |
Method principle and technical path
The molecular biological characteristics of potato yellow dwarf virus are the main basis for the development of this detection method. The detection process includes sample sampling, total RNA extraction, reverse transcription synthesis of cDNA, and PCR amplification and analysis.
- RT-PCR detection: Use specific primers to amplify the target area, and observe specific bands through agarose gel electrophoresis.
- Real-time fluorescence RT-PCR detection: Use fluorescent labeled probe technology to achieve quantitative detection, and judge the results based on the Ct value.
Implementation recommendations and precautions
Actual application case
Suspected PYDV symptoms were detected in a batch of imported potatoes, and the test was carried out according to this standard method:
- Plant materials with virus damage symptoms were extracted as positive controls.
- Total RNA was extracted using Trizol lysis buffer, and cDNA was synthesized by reverse transcription.
- RT-PCR and real-time fluorescence RT-PCR were used for dual detection to ensure the accuracy of the results.
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