GB/T 33120-2016 in English
VALIDQuarantine and Identification of Pear Blister Canker Viroid
- Issued on:2016-10-13
- Implemented on:2017-05-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$214.00
《GB/T 33120-2016梨疱状溃疡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
GB/T 33120-2016 Standard Overview
This standard specifies the quarantine identification method for Pyrus blist spot canker virus (PBCVd), and is applicable to the detection of viruses in imported and exported pear seedlings, tissue culture seedlings and plants.
Background of Standard Formulation
Pyrus blist spot canker virus is a disease that seriously harms pear trees, mainly spread through grafting and machinery. The formulation of this standard aims to standardize the quarantine process and prevent the spread of the virus.
| Dimensions | Contents | Standard requirements |
|---|---|---|
| Scope of application | Pear seedlings, tissue culture seedlings and plants | Objects of entry and exit quarantine |
| Detection method | RT-PCR technology | Molecular hybridization technology |
| Result determination | Positive control verification | Sequence similarity analysis |
Instrument and reagent requirements
The following instruments are required during the detection process:>Ultra-clean bench, PCR instrument, Gel imaging system, etc. The main reagents include RT-PCR reagents and molecular hybridization probes.
Detection process and methods
RT-PCR detection steps
- Nucleic acid extraction: Use specific primers to amplify the target area.
- Reverse transcription and amplification: Synthesize c DNA through the RT step and perform PCR amplification.
- Result determination: Confirm the presence of the virus based on the band size of the amplified product and the sequencing results.
Molecular hybridization detection
- Probe labeling: Use digoxigenin labeling technology.
- Dot blot: Fix the nucleic acid on a nylon membrane for hybridization.
- Signal detection: Use chemiluminescence to display the hybridization results.
Implementation Suggestions
To ensure the accuracy of the test, it is recommended that:
- The laboratory should be equipped with all instruments and equipment required by the standard.
- Operators should receive professional training.
- Establish a complete experimental record and data preservation system.
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