2007年中国四省福氏志贺菌分离菌株的毒力基因检测和PFGE分析

目的分析中国河南等四省的福氏志贺菌分离菌株毒力基因和脉冲场凝胶电泳(PFGE)分型。方法应用PCR方法检测2007年从河南、青海、甘肃和山西分离的262株福氏志贺菌的侵袭性质粒抗原H基因(ipaH)、志贺肠毒素2基因(sen)、志贺肠毒素1基因(set1A)以及侵袭性蛋白基因(ipaBCD)。参考美国CDC的PulseNet实验方法 ,用限制性内切酶NotI对细菌染色体进行酶切,对这些分离菌株进行PFGE分析,使用BioNumerics软件进行聚类分析,并按照PulseNet命名原则对带型进行命名。结果 262株福氏志贺菌分离菌株ipaH、sen、set1A和ipaBCD基因的携带率分别为100%、93.89%、96.18%和92.75%。具有7种毒力基因携带模式,其中89.31%的菌株为Ⅰ型毒力基因携带模式(ipaH+sen+set1A+ipaB-CD+),有99.24%菌株同时携带两种或以上毒力基因。262株福氏志贺菌共分为83个PFGE型别,其中CNJZXN11.0002、CNJZXN11.0003、CNJZXN11.0060、CNJZXN11.0081、CNJZXN11.0137、CNJZXN11.0169、CNJZXN11.0190、CNJZXN11.0195、CNJZXN11.0196、CNJZXN11.0199、CNJZXN11.0217、CNJZXN11.0325和CNJZXN11.0327为13种主要带型。CNJZXN11.0003型菌株在4省均有分布,CNJZXN11.0199为河南菌株的优势带型,CNJZXN11.0137、CNJZXN11.0325和CNJZXN11.0327为山西特有的PFGE带型,与CNJZXN11.0003等主要带型聚类相似性较小。结论四省分离的福氏志贺菌株中ipaH、sen、set1A和iPaBCD毒力基因的携带率较高,而且这些菌株的PFGE型别既具有地区性差异,又具有优势型别的交叉。

Virulence factors detection and pulsed field gel electrophoresis analysis of molecular typing on Shigella flexneri isolated from four provinces in China

ShigellaThe roose of this study was to investigate the virulence genes carrying and molecular eidemiological features of]Shigella flexneri strains isolated from four rovinces of China in 2007.By using CR amlification,the distribution of iaH,Sen,Set1A and iaBCD virulence genes among the 262 strains were 100%,93.89%,96.18% and 92.75%,resectively.All isolates were classified into seven virulence gene carrying tyes.About 89.31% isolates were the virulence gene tyeⅠ（iaH＋sen＋set1A＋iaBCD＋）,while 99.24% strains carried two or more virulence genes.As FGE was erformed according to the laboratory directions for molecular subtying of Shigella by FGE（ulseNet,USA） with some modifications,262 S.flexneri isolates were differentiated into 83 ulse tyes,and CNJZXN11.0002,CNJZXN11.0003,CNJZXN11.060,CNJZXN11.0081,CNJZXN11.0137,CNJZXN11.0169,CNJZXN11.0190,CNJZXN11.0195,CNJZXN11.0196,CNJZXN11.0199,CNJZXN11.0217,CNJZXN11.0325 and CNJZXN11.0327 were the 13 major atterns.CNJZXN11.0003 ulse tye was found in four rovinces,and CNJZXN11.0199 was the revalence ulse tye in Henan.CNJZXN11.0137,CNJZXN11.0325 and CNJZXN11.0327 ulse tyes were found only in Shanxi,and had lower similarity with CNJZXN11.0003.The results roved S.flexneri isolates have a high carrying rate of iaH,Sen,Set1A and iaBCD virulence genes in four rovinces.The distribution of ulse tyes of S.flexneri isolates was different in these regions,but some ulse tyes were crossing distribution.