摘要
本文建立了一种同时检测ＮＤＶ、ＩＢＶ和ＭＧ三种病原体的三重ＰＣＲ技术。根据ＮＤＶ、ＩＢＶ和ＭＧ的基因文库，设计了三对分别与ＮＤＶ、ＩＢＶ和ＭＧ某段基因序列互补的引物，用这三对引物对同一样品中的ＮＤＶ、 ＩＢＶ、 ＭＧ核酸模板进行多重 ＰＣＲ扩增，结果均同时得到了三条特异性大小与实验设计相符的３１０ ｂｐ（ＮＤＶ）、１７２０ ｂｐ（ＩＢＶ）和７３２ ｂｐ（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 １０ ｐｇ的 ＩＢＶ、１ｐｇ的 ＮＤＶ ＲＮＡ模板和 １ｐｇ的 ＭＧ ＤＮＡ模板。
A triplex polymerase chain reaction was optimized to simultaneously detect NDV. IBV and MG. Three sets of specific primers were designed according to the sequences of NDV .IBV and MG at the GeneBank. It was shown the all samples which contained NDV and IBV cDNAs and MG DNA, could be amplified by the triplex PCR using the three sets of primers, yielding three specific bands of NDV 310bp. IBV 1720bp and MG 732bp, but not specific band amplified from other six avian pathogenic virus and bacteria. As little as 10pg of IBV、 1pg of NDV RNA and 1pg of MG DNA was detected using gel electrophoresis in this triplex PCR.
出处
《中国动物检疫》
CAS
2000年第11期20-22,共3页
China Animal Health Inspection
基金
广西留学基金
广西"十百千人才工程"专项资金
关键词
鸡
传染病
NDV
IBV
MG
PCR
病原检测
Triplex Polymerase Chain Reaction Pathogen Detection Newcastle Diseases Virus Infectious Bronchitis Virus Mycoplasma gallisepticumL
