GB/T 34738-2017 in English

VALID

Method of the real-time PCR for the detection of honey bees sacbrood disease

Issued on:2017-11-01
Implemented on:2018-05-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $200.00
$194.00

Standard No:	GB/T 34738-2017
Document status:	VALID
Title in English:	Method of the real-time PCR for the detection of honey bees sacbrood disease
Title in Chinese:	蜜蜂囊状幼虫病荧光PCR检测方法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2017-11-01
Implemented on:	2018-05-01
ICS Classification:	11.220-Veterinary medicine
Professional Classification:	GB-National Standard
Related Keywords:	fluorescent pcr detection method pcr amplification real-time pcr quantitative pcr amplification fluorescent quantitative pcr
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《GB/T 34738-2017蜜蜂囊状幼虫病荧光PCR检测方法》由TC181（全国动物卫生标准化技术委员会）归口，主管部门为农业农村部。

Introduction

Professional interpretation of fluorescent PCR detection method for honey bee sac brood disease

Background of standard formulation and analysis of technical evolution

This standard was proposed by the Ministry of Agriculture of the People's Republic of China and is under the jurisdiction of the National Technical Committee for Animal Health Standardization. Its drafting units include Jilin Entry-Exit Inspection and Quarantine Bureau, Jilin University and Fujian Entry-Exit Inspection and Quarantine Bureau.

The technical basis of the standard is mainly based on fluorescent quantitative PCR (qPCR) detection technology, which is an important progress in the field of molecular biology in recent years and can achieve rapid and highly sensitive pathogen detection.

Standard Framework	Technical Requirements	Scope of Application
Reagents and Materials	The preparation methods of various reagents are clearly specified, including the requirements for key components such as primers and probes.	Applicable to the detection of cystic brood disease in bees and their larvae.
Instruments and Equipment	Requires the use of professional equipment such as fluorescent quantitative PCR amplification instrument and high-speed centrifuge.	Emphasis on the importance of laboratory environment and operating specifications.

Reagent Preparation Instructions

The preparation method of phosphate buffer required by the standard is shown in Appendix A, including the preparation of sodium dihydrogen phosphate and disodium hydrogen phosphate solutions and high-pressure sterilization. The preparation of diethyl pyrocarbonate treated water requires standing and high-temperature sterilization.

The specific preparation method of RNA extraction reagent and preservation solution is: add chloroform to the total RNA extraction reagent lysis solution, mix well, centrifuge and take the supernatant for use.

Overview of the detection process

Sample collection and processing:According to the requirements of GB/T18088, collect 20 to 30 suspected infected honey bee larvae.
RNA extraction:Use total RNA extraction reagent for lysis, and take the supernatant after centrifugation for subsequent testing.
cDNA synthesis:Completed on an ice box, using random primers and reverse transcriptase to catalyze the generation of cDNA.
PCR amplification and analysis:Prepare the PCR reaction mixture, add the fluorescent probe and perform quantitative PCR amplification, and set appropriate cycling conditions to detect the amplification amount of the target virus.

More samples: JJG 21-2008 JGJ 94-2008 GB 18306-2015

Sample only — not a preview of GB/T 34738-2017

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