摘要
本文建立了一种同时检测NDV、IBV、ILTV和MG 4种病原体的多重PCR技术。根据NDV、IBV、ILTV和MG的基因文库 ,设计了 4对分别与NDV、IBV、ILTV和MG某段基因序列互补的引物 ,用这 4对引物对同一样品中的NDV、IBV、ILTV、MGRVA或DNA模板进行多重PCR扩增 ,结果均同时得到了 4条特异性大小与实验设计相符的 310bp(NDV)、172 0bp(IBV)、6 47bp(ILTV)、和 732bp(MG)多重PCR扩增带 ,而对其他 6种禽病病原的PCR扩增结果均为阴性 ;敏感性测定结果表明 ,该多重PCR技术能检出 10pg的IBV、1pg的NDVRNA模板和 10pg的ILTV、1pg的MGDNA模板。
A multiplex polymerase chain reaction was optimized to simultaneously detect four pathogens of NDV,IBV,ILTV and MG in this article.Four sets of specific primers were designed according to the sequences of NDV,IBV,ILTV and MG in the genebank.It was showed that all samples which contained NDV,IBV cDNA and ILTV,MG DNA,could be amplified by the multiplex PCR using these four sets primers,yielding four specific bands of NDV 310bp,IBV 1720bp,ILTV647bp and MG 732bp.But not specific band amplified from other six avian pathogenic viruses and bacterium.As little as 10pg of IBV,1pg of NDV RNA and 10pg of ILTV,1pg of MG DNA were detected using gelelectrophoresis in this multiplex PCR.
出处
《中国预防兽医学报》
CAS
CSCD
2000年第6期443-446,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
广西留学基金
国家非教育系统留学回国人员择优基金的项目
关键词
多重聚合酶链式反应
检测
鸡新城疫病毒
鸡传染性支气管炎病毒
鸡传染性喉气管炎病毒
鸡毒支原体
Multiplex Polymerase Chain Reaction
Detection
Newcastle diseases virus
Infectious bronchitis virus
Infectious laryngotracheitis virus
Mycoplasma gallisepticum
