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目的对宁波地区鼠类携带的钩端螺旋体进行聚合酶链反应(PCR)检测并分析基因序列特征。方法设计钩端螺旋体的sec Y基因和23S r DNA基因引物,对宁波梅山口岸2014年5月-12月捕获的鼠类样本进行PCR检测及核苷酸序列测定与系统进化分析。结果共检测72份鼠类样本,其中5份黑线姬鼠经检测为阳性,分离获得钩端螺旋体株DXLP1501,该菌株sec Y基因扩增片段大小为285 bp,23S r DNA基因扩增片段大小为484 bp。同源性比对结果显示,23S r DNA基因片段的系统发生分析显示DXLP1501与博氏螺旋体株L550位于同一分支,遗传距离为0.005;sec Y基因片段的系统发生分析显示DXLP1501与5株博氏钩端螺旋体菌株和1株梅氏钩端螺旋体位于同一分支,遗传距离为0.011。结论本研究证实了宁波地区鼠类中存在博氏钩端螺旋体的流行。
Objective To detect and analyze the sequence characteristics of the leptospira carried by rodents in Ningbo by polymerase chain reaction (PCR). Methods The sec Y gene and the 23S r DNA gene primers of Leptospira were designed and used to detect the PCR products, nucleotide sequences and phylogenetic analysis of murine samples captured from May to December in Ningbo Meishan Port. Results A total of 72 mouse samples were detected. Among them, 5 Apodemus agrarius strains were tested positive for DXLP1501. The sec Y gene fragment was 285 bp in length and 23S r DNA fragment was amplified Is 484 bp. Phylogenetic analysis showed that the phylogenetic analysis of 23S r DNA fragment showed that DXLP1501 was located on the same branch as B. lamblia strain L550 with a genetic distance of 0.005. Phylogenetic analysis of sec Y gene fragment showed that DXLP1501 and 5 strains of BorgH Leptospira strains and a strain of Leptospira interrogans were located in the same branch with a genetic distance of 0.011. Conclusion This study confirms the prevalence of Leptospira borgpeters in the rodents in Ningbo area.