摘要
利用逆转录－ＰＣＲ（ＲＴ－ＰＣＲ）特异性扩增鸡传染性支气管炎病毒（ＩＢＶ）基因组中Ｍ基因和Ｎ基因之间一段核酸片段。以ｐＵＣ１９质粒载体将此片段克隆，用ＥｃｏＲＩ和ＨｉｎｄⅢ酶切此重组质粒，回收克隆片段后，制成生物素标记的核酸探针。用ＲＴ－ＰＣＲ及生物素核酸探针法分别对ＩＢＶ，ＩＢＤＶ，ＩＬＴＶ，ＭＤＶ及ＮＤＶ进行检测，结果证明该方法为ＩＢＶ特异性检测方法。对人工感染ＩＢＶ的ＳＰＦ鸡口腔棉拭样品进行跟踪检测，证明本方法能在ＳＰＦ鸡接毒后１～１０ｄ内检出ＩＢＶ。
Reverse transcription of viral RNA and polymerase chain reaction were used to amplify a cDNA fragment between M gene and N gene of infectious bronchitis virus (IBV). The amplified DNA fragment was cloned into pUC19. The recombinant plasmid was digested by Hind Ⅲ and Eco RI to purify the cloned DNA fragment. The purified DNA fragment was labelled by biotin as a DNA probe. The dot hybridization assay with the DNA probe was used to detect IBV, NDV, ILTV and IBDV. Only IBV showed positive. The PCR and DNA probe have been used to detect tracheal swab samples from SPF chicken which were challenged by IBV. It was shown that the PCR and DNA probe assay can detect IBV from the chicken from 1 to 10 days after being challenged.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1997年第5期431-433,共3页
Chinese Journal of Veterinary Science
基金
国家"八五"科技攻关项目
