布鲁氏菌病VirB8-PCR诊断试剂盒的特异性评价

Sequence Analysis on VirB8 genes of 7 Different brucella

使用VirB8-PCR诊断试剂盒对布鲁氏菌标准株、疫苗株及新疆分离株共7株布鲁氏菌的VirB8基因序列扩增、克隆并与Genbank中发表的VirB8基因序列(AF226278)进行比较和分析,发现:该基因非常保守,7株布鲁氏菌的同源性在99.2%以上,其中A菌与Rev1基因序列100%同源,B菌与Genbank发表的基因序列100%同源;牛种(544A、A19)和羊种(M5、Rev1)分别各自在相同部位发生了相同的碱基突变,因此VirB8基因有可能做为布鲁氏菌疫苗菌的鉴定分类基因。VirB8基因在布鲁氏菌中高度保守,可作为布鲁氏菌检测模板,VirB8-PCR诊断试剂盒特异、敏感,检测结果可靠。

Using VirB8-PCR of brucella to amply VirB8 fragment, and then analysis the sequence of VirB8 genes of 7 different brucellas. A pair of primers were designed for amplification of brucella VirB8 genes according to the complete genomic sequence blished in GenBank. VirB8 genes were amplified from 11 different bucellas by polymerase chain reaction（VirB8-PCR）. The PCR products were cloned in pMD18-T vector for sequencing. Results Sequence analysis showed that the autoploidy of VirB8 genes of 7 different brucellas is 99.2%. Sequence analysis on VirB8 genes of 7 different Brucellas showed that brucella VirB8 genes were highly conserved and the kit is available.