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鉴别新城疫强、弱毒一步RT-PCR方法的建立及应用

Differentiation of velogenic and lentogenic strains of NDV by one step RT-PCR

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【作者】 明文龙尹仁福谢光耀薛聪王珂袁乾亮丁壮

【Author】 MING Wen-long;YIN Ren-fu;XIE Guang-yao;XUE Cong;WANG Ke;YUAN Qian-liang;DING Zhuang;College of Veterinary Medicine,Jilin University;

【机构】 吉林大学动物医学学院

【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。

【Abstract】 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.

【关键词】 NDV一步法RT-PCRF基因鉴别诊断
【Key words】 Newcastle disease virusone step RT-PCRF genedifferentiation
【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)
  • 【文献出处】 中国兽医学报 ,Chinese Journal of Veterinary Science , 编辑部邮箱 ,2015年07期
  • 【分类号】S852.65
  • 【被引频次】9
  • 【下载频次】242
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