30株鸡传染性支气管炎病毒标准株与分离株S1基因的克隆及序列分析

Cloning and nucleotide sequencing analysis of the S1 gene from 30 infectious bronchitis virus strains with standard reference and isolate strains

利用IBVS1基因特异性引物对30株IBV毒株进行RT-PCR扩增,产物经纯化后进行测序,并利用DNAstar MegAlign软件分析S1基因核苷酸序列及推导的氨基酸序列。结果表明,广西分离株GXIB/02与Mass41、Conn、Ark、PA、Del、JMK、H52参考株的核苷酸序列同源性和氨基酸序列同源性均较低,与美国80年代分离的毒株MW34、hotle、cal15、AustT株的序列同源性也很低。这一结果与血清学试验结果相吻合,说明广西分离株GXIB/02与当今流行的标准株以及过去流行的毒株都不相同,是一个新的变异株,由此推断广西已有不同血清型IBV毒株存在,可能导致新的变异株流行。

Based on the published IBV gene sequences,a pair of specific primers was designed and synthesized for amplification of the S1 genes IBV strains. Total 30 of RNA extractor from IBV strains including reference and field isolates strains were amplified by reverse transcription and polymerase chain reaction （RT-PCR）. The PCR products were purified and then sequenced. S1 gene nucleotides and predicted amino acids were analyzed using DNAstar MegAlign software. The results revealed that the isolate strain GXIB/02 had less than 80% similarity homology with the 7 standard reference strains Mass 41, Conn, Ark, PA, Del, JMK, H52 and domestic strains MW34, hotle, Cal 15 isolated from America in 1980s. The serum antibody neutralization test and nucleotide sequencing analysis results demonstrated that GXIB/02 strain isolated from Guangxi was a new variant IBV in China.