摘要
设计2对引物分别用于新城疫强、弱毒的检测。使用强毒引物F1、R1可从NDV强毒株中特异性扩增出349bp的目的片段,使用弱毒引物F2、R2可从NDV弱毒株中特异性扩增出255bp目的片段,该方法对H9亚型禽流感病毒(AIV)、传染性支气管炎病毒(IBV)和传染性法氏囊病毒(IBDV)的检测结果均为阴性。灵敏性试验结果显示,该方法对NDV强、弱毒株的最小检出量分别为1pg和10pg。利用该方法对7份临床样品进行检测,结果与测序结果一致,说明该方法可用于新城疫强、弱毒的快速鉴别诊断。
Two pairs of primers for differentiation of velogenic and lentogenic strains of NDV were designed based on the difference of the nucleotide sequence at F protein cleavage site at 112 within NDV genome.A single PCR band of 349 bp was obtained only from velogenic,not lentogenic strains RNA when used the primer pair F1 and R1,while a single 255 bp band was obtained only from lentogenic strains RNA when used the primer pair F2 and R2.Furthermore either 349 bp or255bp was detected from RNA of avian influenza virus（AIV）H9subtype,infectious bronchitis virus（IBV）and infectious bursaldisease virus（IBDV）.Sensitivity of the two pairs primers was 1pg and 10 pg,respectively.Application of one-step RT-PCR to 7clinical NDV samples demonstrated the potential for identifing the velogenic and lentogenic strains of NDV.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2015年第7期1056-1059,共4页
Chinese Journal of Veterinary Science
基金
国家公益性行业(农业)科研专项(201303033)
国家自然科学基金资助项目(31272561
31472195
31402195)
吉林大学大学生国家级创新基金项目(2014A81365)
