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目的对安徽省2008-2010年宋内氏志贺氏菌进行分子流行病学分析,为安徽省的菌痢检测和监测搭建分子平台。方法应用纸片扩散(kirby-bauer,K-B)法进行药物敏感试验;应用聚合酶链式反应(polymerase chain reaction,PCR)方法对20株宋内氏志贺氏菌进行基因分析;运用脉冲场凝胶电泳(pulsed-field gel electrophoresis,PFGE)技术进行亲缘性分析。结果 20株宋内氏志贺菌对氟哌酸(ciprofloxacin,CIP)和先锋必(cefoperazone,CPZ)最为敏感,敏感率达100%;多重PCR结果显示除志贺毒素1(shiga toxin 1,SET1)为阴性外,其余三种基因:侵袭性质粒H抗原(invasion plasmid antigen H,ipaH)、志贺肠毒素2(shiga toxin 2,SET2)、志贺菌增殖和侵袭相关的毒力基因(invasion associated locus,ial)皆为阳性;20株菌用XbaⅠ酶切后经PFGE电泳后可分为6个主要型别。结论安徽省受试20株宋内氏志贺菌多重耐药现象严重且基因型高度一致;菌型呈多种型别并存的趋势,说明安徽省宋内氏菌的来源属于多克隆系。
Objective To analyze the molecular epidemiology of Shigella sonnei from 2008 to 2010 in Anhui province and establish a molecular platform for the detection and monitoring of bacillary dysentery in Anhui Province. Methods The kirby-bauer method was used for drug susceptibility testing. 20 strains of Shigella sonnei were analyzed by polymerase chain reaction (PCR) Gel electrophoresis (pulsed-field gel electrophoresis, PFGE) technology for affinity analysis. Results 20 strains of Shigella flexneri were most sensitive to ciprofloxacin (CIP) and cefoperazone (CPZ), with a sensitivity of 100%. Multiplex PCR showed that shiga toxin 1 (SET1 ) Were negative, the other three genes: invasion plasmid antigen H (ipaH), shiga toxin 2 (SET2), Shigella proliferation and invasion-related virulence genes (invasion associated locus, ial) were all positive; 20 strains were digested with Xba Ⅰ and separated by PFGE electrophoresis into 6 major types. Conclusion The multi-drug resistance of 20 strains of Shigella sonnei in Anhui province was severe and the genotypes were highly consistent. The trend of multiple types of coexisting bacteria was observed, indicating that the origin of the genus Clostridium in Anhui Province belongs to the polyclonal system.