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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。
Objective To analyze the virulence genes and pulsed-field gel electrophoresis (PFGE) typing of Shigella flexneri isolated in four provinces of Henan, China. Methods 262 strains of Shigella flexneri isolated from Henan, Qinghai, Gansu and Shanxi in 2007 were genotyped by PCR, including ipaH, ssh, Gene (set1A) and invasive protein gene (ipaBCD). Reference to the CDC’s PulseNet experimental method in the United States, digestion of bacterial chromosomes with restriction endonuclease NotI, PFGE analysis of these isolates, cluster analysis using BioNumerics software and naming of bands according to the PulseNet nomenclature. Results The carrying rates of ipaH, sen, set1A and ipaBCD genes of 262 strains isolated from Shigella flexneri were 100%, 93.89%, 96.18% and 92.75% respectively. There were 7 virulence genes carrying patterns, of which 89.31% strains were type I virulence genes carrying mode (ipaH + sen + set1A + ipaB-CD +), 99.24% strains carrying two or more virulence genes simultaneously. 262 strains of Shigella flexneri were divided into 83 PFGE types, of which CNJZXN11.0002, CNJZXN11.0003, CNJZXN11.0060, CNJZXN11.0081, CNJZXN11.0137, CNJZXN11.0169, CNJZXN11.0190, CNJZXN11.0195, CNJZXN11 .0196, CNJZXN11.0199, CNJZXN11.0217, CNJZXN11.0325 and CNJZXN11.0327 are the 13 main belt types. CNJZXN11.0003 strains are all distributed in 4 provinces, CNJZXN11.0199 is the dominant belt type of Henan strains, CNJZXN11.0137, CNJZXN11.0325 and CNJZXN11.0327 are endemic PFGE types in Shanxi, and CNJZXN11.0003 and other major types Clustering is less similar. CONCLUSIONS The ipaH, sen, set1A and iPaBCD virulence genes isolated from four strains of Shigella flexneri isolated from four provinces have high carriage rates. Moreover, the PFGE types of these isolates have both regional differences and dominant type crosses.