摘要
根据国外报道的鸡传染性支气管炎病（ＩＢＶ）Ｇｒａｙ株的纤突蛋白（Ｓ１）核苷酸序列，自行设计合成了Ｓ１蛋白主要抗原决定簇基因两侧的１对引物，跨幅为１．０ｋｂ。用该引物对从内蒙古、天津、山东等地分离的３株ＩＢＶ进行ＲＮＡ提取，反转录后ＰＣＲ，经琼脂糖凝胶电泳结果表明，３株毒株均获得了与预期一致的ＰＣＲ产物；此外，ＩＢＶＰＣＲ灵敏度的测定结果表明，ＰＣＲ方法可检测出１０ｐｇ的模板，ＰＣＲ酶切结果为，ＨａｅⅢ、ＴａｇⅠ酶对３株ＰＣＲ产物均有１个酶切位点。
A pair of specific primers were designed and synthesized according to the nucleotide sequence of S 1 protein gene form Gray strain of infectious bronchtis virus.The cDNAs of S 1 protein gene of IBV Inner Mongolia、Tianjin and Shandong isoloates were amplified by reverse transcription polymerase chain reaction(RT-PCR).Results show that we have obtained 1.0kb expected size cDNA fragments from the RNAs of these three isolates.The limit of IBV RNA detection by RT-PCR assay was 10pg of RNA in the RT reaction.Results of restriction mapping analysis show that on one of the cDNA fragments from three isolates there were one HaeⅢ and TaqⅠ sites and there was no MspⅠsite.
出处
《中国兽医杂志》
CAS
北大核心
1997年第1期5-7,共3页
Chinese Journal of Veterinary Medicine
