GB/T 40135-2021 in English

VALID

Detection and identification of Xylophilus ampelinus (Panagopoulos) Willems et al.

Issued on:2021-05-21
Implemented on:2021-12-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $250.00
$243.00

Standard No:	GB/T 40135-2021
Document status:	VALID
Title in English:	Detection and identification of Xylophilus ampelinus (Panagopoulos) Willems et al.
Title in Chinese:	葡萄细菌性疫病菌检疫鉴定方法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2021-05-21
Implemented on:	2021-12-01
ICS Classification:	65.020.01-Farming and forestry in general
Chinese Classification:	B16-Plant quarantine, pest control
Professional Classification:	GB-National Standard
Related Keywords:	molecular detection detection sensitivity elisa detection systematic detection method specific fragment pcr detection target comparison
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《GB/T 40135-2021葡萄细菌性疫病菌检疫鉴定方法》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

Analysis of the core content of the standard

This standard specifies the systematic detection method for grape bacterial blight (Xylophilus ampelinus), covering the technical specifications of the entire process including field sampling, laboratory isolation, immunological testing and molecular biological identification.

Pathogen characteristics and hazards

Characteristic dimensions	Typical manifestations	Detection significance
Morphological characteristics	Rod-shaped, monopolar flagellum, Gram-negative	Basis for initial screening under microscope
Culture characteristics	YPGA culture medium forms 2mm light yellow colonies in 7-12 days	Isolation and culture judgment criteria
Molecular markers	ITS sequence/16s-23s rDNA specific fragment	PCR detection target

Comparison of detection technologies

Selection of immunological and molecular detection schemes

The standard recommends the dual method validation principle:

ELISA detection: suitable for large-scale initial screening, easy to operate but requires antibody specificity validation
PCR technology: includes three schemes: ordinary PCR (129bp), nested PCR (277bp) and real-time fluorescence PCR (91bp), and the detection sensitivity increases successively

Typical case shows: For asymptomatic samples, the nested PCR + pathogenicity experiment combination scheme is recommended.

Key points for the implementation of the standard

Warnings for key operations

Sample processing must strictly follow the principle of sampling at the borderline of disease and health
The phenol-chloroform purification step cannot be omitted during DNA extraction
The adaptability of YPGA culture medium to pH 6.5-7.0 needs to be verified

Analysis of technological evolution

Compared with historical methods, this standard has three major breakthroughs:

Real-time fluorescence PCR technology is introduced (Ct value determination standard)
The gradient dilution separation method for asymptomatic materials is standardized
The concentration parameters for glycerol storage (20-30%) are clarified

According to literature tracing, the detection sensitivity has been improved from 10^3CFU/mL of traditional culture method to 10^1CFU/mL of molecular detection.

More samples: QC/T 942-2021 JC/T 2453-2018 JJG 21-2008

Sample only — not a preview of GB/T 40135-2021

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