霍乱弧菌O1群毒力基因分布和DNA分子特征  

DNA characterization and the virulence gene distribution of Vibrio cholerae O1

作　　者：段广才[1,2] 高守一[1,2] 刘延清[1,2] 祁国明[1,2] 刘彩莲[1,2] 

机构地区：[1]河南医科大学公共卫生学院流行病学教研室 [2]中国预防医学科学院流行病学微生物学研究所

出　　处：《河南医科大学学报》1998年第3期35-40,共6页Journal of Henan Medical University

基　　金：国家自然科学基金

摘　　要：由于霍乱弧菌（ＶＣ）Ｏ１群菌株中仅有部分可以引起霍乱流行，研究其主要毒力基因分布和ＤＮＡ分子特征，已成为目前学者们关注的重要课题。应用原位杂交法对４７１株Ｏ１群ＶＣ的主要毒力基因分布情况进行检测，用核酸脉冲场凝胶电泳对其中３４株进行染色体ＤＮＡ特征和ＣＴ基因Ｓｏｕｔｈｅｒｎ杂交分析。结果：在ＳａｌＩ酶切图谱中，所有埃尔托流行株（ＥＳＥＶＣ）具有一致的酶切图谱；古典型菌株（ＣＶＣ）除１１１９菌与ＥＳＥＶＣ图谱相同外，其他均具有一致的特征性图谱。ＣＶＣ和ＥＳＥＶＣ之间有较微小的差异。埃尔托非流行株（ＮＳＥＶＣ）的图谱与ＥＳＥＶＣ之间有较大差异。在ＳｍａＩ和ＸｂａＩ酶切图谱中，结果与上述一致，其中ＣＴ基因缺失的ＥＳＥＶＣ菌株７７４３和８０１２７５具有较相似的ＳｍａＩ酶谱。用ｃｔｘ、ｚｏｔ、ａｃｅ、ＲＳ１为探针，对４７１株Ｏ１群霍乱弧菌进行毒力基因检测；结果：ＣＶＣ菌株全部阳性；ＥＳＥＶＣ菌株中，上述基因阳性率分别为：９７．３％、９７．５％、９７．３％、９９％；在１０株ＣＴ基因缺失的ＥＳＥＶＣ菌株中，有三株为ｚｏｔ基因阳性；在ＮＳＥＶＣ菌株中，有一株为ｚｏｔ和ＲＳ１阳性。美国海湾分离的６株产毒ＶＣ上述基因阳性。结果?Vibrio cholerae O1 (VC O1) are considered the pathogens of epidemics and pandemics of cholera. Distiguishing the different strains of VC O1 on molecular genetics is the key work for cholera prevention and control. Here we reported the results of the distributions of the virulence genes (ctx, zot, ace, RS1) among 471 isolates of VC O1 by colony hybridization, and the genomic DNA characteristics of 34 isolates of VC O1 by PFGE and Southern blot with ctx probe. The results showed that with SalI digestion, all of patterns of epidemigenic strains of El Tor (ESEVC) were identical, and all of the patterns of Classical Vibrio cholerae (CVC) (excluding 1119 similar to ESEVC) were identical. There some little differences between CVC and ESEVC and obvious differences between ESEVC and NSEVC (non epidemigenic strains of El Tor). In the SmaI and XbaI patterns the results were similar with those above. By the colony hybridization with the virulence gene brobes of ctx, zot, ace, and RS1, all of CVC were positive. Among 364 isotlates of ESEVC, the positive proportions for these gene probes were 97.3%, 97.5%, 97.3%, and 99%. Among 75 isolates of NSEVC there was only one isolate positive for zot and RS1.

关 键 词：O1群 霍乱弧菌 毒力基因分布 DNA 分子特征 

分 类 号：R378.3[医药卫生—病原生物学]