福建农业学报2018，Vol.33Issue(3)：219-224,6.DOI:10.19303/j.issn.1008-0384.2018.03.001
鸭肠炎病毒强毒和疫苗弱毒鉴别诊断PCR方法的建立
Rapid PCR Method for Differentiating Virulent and Vaccine Enteritis Viruses in Ducks
摘要
Abstract
To accurately and rapidly differentiate between the virulent and the attenuated vaccine strains of duck enteritis viruses (DEVs),sequences of the virus UL2 genes were retrieved from the databank at GenBank for the study.A 528 bp continuous nucleotide deletion region was found on the genes of both strains.Using the primer design software,Oligo 7.0,specific primers targeting for the development and optimization of the PCR differentiation diagnosis methodology based on the UL2 characterization were selected.The optimal annealing temperature was determined to be 55℃ rendering amplified segments for the DEV virulent and the vaccine strains at 1019 bp and 491 bp,respectively.The established protocol was sensitive with a low detection limit of 15.3 pg;and,specific without cross amplification with common duck origin pathogens,such as,Muscovy duck parvovirus,duck circovirus,goose parvovirus,Escherichia coli,Rimerella anatipstifer and Pasteurella multocida.Consequently,it was considered applicable for future studies on the pathogenic mechanism of DEV.关键词
鸭肠炎病毒/强毒/疫苗弱毒/UL2基因/PCR/鉴别诊断Key words
duck enteritis virus/virulent strain/attenuated vaccine strain/UL2 gene/PCR/differential diagnosis分类
农业科技引用本文复制引用
万春和,刘荣昌,陈翠腾,程龙飞,傅光华,施少华,陈红梅,傅秋玲,黄瑜..鸭肠炎病毒强毒和疫苗弱毒鉴别诊断PCR方法的建立[J].福建农业学报,2018,33(3):219-224,6.基金项目
国家水禽产业技术体系建设专项(CARS-42) （CARS-42）
国家自然科学基金项目(31602068) （31602068）
福建省财政专项——福建省农业科学院青年科技创新团队建设项目(STIT2017-3-10) （STIT2017-3-10）
福建省农业科学院青年科技英才项目(YC2015-12) （YC2015-12）
福建省科技计划项目——省属公益类科研院所基本科研专项(2018R1023-5、2017R1023-7) （2018R1023-5、2017R1023-7）
