[Objective] To establish a simple and rapid molecular typing method tor Vibrio parahaemolyticus carrying and non-carrying virulence genes based on molecular motor.[Methods] Four probes specific to virulence genes tdh and trh,species-specific genes tlh and toxR of V.parahaemolyticus were synthesized,and four molecular motor biosensors were constructed by connecting probes to F0F1-ATPase molecular motors through biotin-streptavidin system,respectively.Ten strains of V.parahaemolyticus were classified by the biosensors,and the results were compared with PCR-Electrophoresis-Gel imaging results.Further more,the detection sensitivities and specificities of the molecular motor biosensors were studied.[Results] There were ten strains carrying tdh and none carrying trh,while all ten strains carry tlh and toxR,which was consisitent with the results of PCR-Electrophoresis-Gel imaging.The detection limits of molecular motor biosensors for tlh,toxR,tdh and trh were estimated to be 1 pg/reaction system,and the detection limits of PCR-Electrophoresis-Gel imaging for tlh,toxR,tdh and trh were estimated to be 10 pg/reaction system.The molecular motor biosensors could recognize tlh,toxR,tdh and trh of V.parahaemolyticus specifically.[Conclusion] A molecular typing method was constructed based on molecular motor biosensors and was used to diagnose the pathogenicity of V.parahaemolyticus rapidly and specifically.The detection limits was 10 times higher than those of PCR-Electrophoresis-Gel imaging.The method is easy,rapid,time-saving and labor-saving,especially suitable for the basic laboratories of CDC and port quarantine departments to perform suiveillance and epidemiological traceability of cholera.%[目的]建立基于分子马达技术的简便快速的分子分型方法,对携带和非携带毒力基因的副溶血性弧菌进行快速分类.[方法]以F0F1-ATPase为核心构建分子马达,以副溶血性弧菌毒力基因tdh、trh和种特异性基因tlh、toxR为靶基因设计4个探针.通过生物素-亲和素系统将探针与分子马达连接构建F0F1-ATPase分子马达生物传感器,对10株副溶血性弧菌分离株进行分类,并与PCR-电泳-凝胶成像结果进行比较;同时对生物传感器的检测灵敏度和特异性进行研究.[结果]10株试验菌株中10株tdh阳性,0株trh阳性,而10株菌都携带tlh和toxR,与PCR-电泳-凝胶成像结果一致；分子马达生物传感器的最低检测限为1 pg/反应体系,且能够对副溶血性弧菌特异性识别,PCR-电泳-凝胶成像方法的最低检测限为10 pg/PCR反应体系.[结论]建立了基于分子马达的分子分型方法,能够对副溶血性弧菌的致病性进行快速诊断,检测灵敏度比PCR-电泳-凝胶成像方法高了10倍,而且特异性非常高.该方法简便、快速、省时、省力,适用于地方疾控部门和口岸检疫部门的基层实验室开展副溶血性弧菌监测和流行病学溯源工作.
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