摘要
目的 建立产单核细胞李斯特菌的分子检测方法及其亚分型体系。方法 对LM菌株进行hly基因的PCR检测 ,并扩增其中 10株PCR检测阳性菌株的actA基因 3’末端的 82 7bp的DNA片段 ,对扩增的片段进行测序。利用遗传进化树分析软件分析。结果 2 4株LM的hly基因扩增均出现 85 0bp左右特异性片段 ,而非LM株未出现该特异性片段。所建立的分子亚分型方法将 10株LM归为两个遗传谱系。结论 新建立的PCR法检测LM具有较高的灵敏性与特异性 ,LM分子亚分型体系的建立为进一步研究不同LM分离株的群体遗传学、流行病学及生态学奠定了技术基础。
Aim To establish the method for identifying and subtyping the isolates of Listeria monocytogenes.Method The specific hly,actA genes of Listeria monocytogenes isolates were amplified respectively.The amplified fragments of actA gene were sequenced and then analyzed with MEGA2 version software,and then the phylogenetic tree of Listeria monocytogenes was established.Result The 850bp fragment of amplified hly gene fragment was yielded in all Listeria monocytogenes,while in non- Listeria monocytogenes,this specific fragment didn't occur.These Listeria monocytogenes could be divided into two different lineages with this DNA-based subtyping technique.Conclusion The hly gene-based PCR method for identifying Listeria monocytogenes yielded high specificity and sensitivity.The establishment of DNA-based subtyping method provided new insight for studying the population genetics,ecology and epidemiology of Listeria monocytogenes.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2003年第5期44-47,共4页
Chinese Journal of Zoonoses
基金
教育部"高校青年教师奖"资助 ( 175 )
