荧光定量PCR和DNA分子分型在菌痢暴发中的应用

目的]探讨实时荧光PCR快速检测和脉冲肠凝胶电泳（PFGE）分型方法在菌痢暴发流行病学研究中的应用。方法]采用实时荧光PCR检测技术对一起菌痢患者的24份肛拭增菌液进行快速检测，同时从患者肛拭中分离的5株福氏4型志贺氏菌采用脉冲场凝脉电泳的方法进行了分子分型。结果]在这一起菌痢疫情中，实时荧光PCR快速检测24份肛拭增菌液．5份阳性，并从PCR阳性的增菌液中分离到5株福氏4型志贺氏菌，而且5株志贺氏菌的脉冲场凝胶（PFGE）图谱显示DNA条带完全一致，具有高度的同源性。结论]实时荧光定量PCR技术特异性强，灵敏度高，能在采样后数小时内检测到病原菌的DNA，达到快速诊断的目的。PFGE具有分型能力强，重复性好等特点，能直观地判断致病菌的亲缘关系，及时查明暴发流行的传染源从而有效控制疫情的蔓延，在志贺氏菌的分子流行病学研究中具有重要作用。

THE APPLICATION OF REAL-TIME QUANTITATIVE PCR AND DNA MOLECULAR TYPING ON DEALING WITH THE OUTBREAK OF SHIGELLOSIS

Objective]To explore the application of Real-time PCR procedure and DNA molecular typing method on dealing with the outbreak of shigellosis.Methods]Real-time PCR procedure was used to rapidly detect shigellosis in 24 shares enrichment broth of anus mops from people with dysentery,and 5 isolates of Sh.flexneri from mops were typed by pulsed-field gel electrophoresis(PFGE)simultaneously.Results]5 shares were shown positive from the 24 shares of enrichment broth of anus mops by Real-time quantitative PCR procedure in this outbreak incidence,and then 5 isolates of Sh.flexneri were isolated from the enrichment broth.The PFGE patterns of the 5 strains were identical,which expressed the high homology of them.Conclusion]Real-time PCR procedure is a method with high sensitivity,specificity.It can detect the DNA of the bacteria in few hours,which make the diagnosis rapid.The fine typing ability and reliability of PFGE is helpful for finding out the heredity background of intestinal pathogen and the infectious resource of the outbreak to effectively prevent the epidemic situation from speading.It is important in the molecule epidemiological research.