人感染猪链球菌病的病原分离与PCR方法的快速检测.pdfVIP

实用预防医学 2008年2月 第 15卷 第1期 Practical Preventive Medicine，Feb．2008，Vol 15，No．1 209 文章编号：1006—3110(2008)01—0209—03 人感染猪链球菌病的病原分离与PCR方法的快速检测 韩丽娟 ，曾晓燕 ，李 莉 ，张学军 ，杨广泽 ，朱磷扬 摘要： 目的 研究人感染猪链球菌的生物学特征，并建立一种快速、特异且敏感的检测方法。 方法 将疑似猪链 球菌感染患者的血液进行增菌培养，对所分离的病原菌进行染色形态观察、AP1 32 Strep生化鉴定；同时从该疑似患者血液 (抗凝血)中提取DNA为模板，并合成引物检测猪链球菌特异性 16SrRNA基因片段、2型猪链球菌特异性荚膜多糖编码基 因片段cps2J、溶菌酶释放相关蛋白编码基因片段mrp、细胞外蛋白因子编码基因片段 ef。 结果 PCR扩增结果证实是 猪链球菌2型，而从血液中分离的病原菌，经AP1 32 Strep生化鉴定，也证实为猪链球菌2型。 结论 人感染猪链球菌的 病原学及PCR检测符合猪链球菌2型，同时该PCR反应体系是一种检测和鉴定猪链球菌2型的快速、特异且敏感的方法。 关键词：猪链球菌；生化鉴定；PCR；基因 中图分类号：R378 文献标识码：A Application of Pathogen Isolation and PCR Assay for Rapid Detection of Streptococcus Suis Serotype2 from Human Case HANLi—juan‘，ZENGXiao—jan ，LJLi‘，et a1．(1．LianyungangMunicipalCenterforDiseaseControl andPrevention， Lianyungang 222003，Jiangsu，China； 2．Jiangsu Provincial Center for Disease Control and Prevention，Nanjing 210009， Jiangsu，China) Abstract： Objective To isolate StreDtococcus suis strains and to study the biological characteristics，a highly specif— ic，sensitive and rapid method for the detection of streptogoccus suis by PCR was developed． Metbods Blood—agar sub— strates were used to cultivate and isolate pathogens from the blood(anticoagulation)of suspect patients．The morphology of the isolated bacteria was observed after staining and the specific serotypes were identified by API 32 Strep biochemical tests． The blood sample(anticoagulation)was taken from the suspect patients，DNA was extracted as template and the primer was synthesized according to the gene sequence to detect 16SrRNA，capsule polysaccharide 2J(cps2J)，muramidase—released protein(mrp)and extracellular factor protein(EF)． Results Streptococcus suis serotype 2 was founded in the specimens frOm the suspect patients by PCR and was further confirmed by API 32 Strep