天然水生环境中霍乱弧菌种特异性快速鉴定及毒力测定方法

B. Chakraborty, K. Zaman, Majibur Rahman
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引用次数: 5

摘要

由产毒霍乱弧菌引起的霍乱是发展中国家面临的一个主要公共卫生问题,在这些国家,霍乱的爆发经常季节性发生,并特别与贫穷和卫生条件差有关。人们普遍认为,自1817年第一次霍乱大流行爆发以来,已经发生了七次不同的霍乱大流行。霍乱弧菌能够在水生环境中存活较长时间,被认为是河口和咸淡水中的原生物种。因此,本研究旨在从自然环境样品中分离霍乱弧菌,然后通过常规和分子生物学技术鉴定。根据初步鉴定结果,随机纳入10株分离菌。经血清学检测(玻片凝集法)确定分离株血清型,经PCR法再次确认的O1、O139和非O1/O139血清型分别为3、2和5株。最后,采用多重PCR方法分析分离株的毒力,发现5株分离株含有霍乱弧菌主要毒力因子ctx基因。关键词:霍乱弧菌,单纯PCR,多重PCR,血清型,毒力Â doi:10.3329/sjps.v1i1.1811 S. J. Pharm。科学通报1(1&2):69-75
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Rapid Method for Species-Specific Identification and Determination of Toxigenicity of Vibrio Cholerae from Natural Aquatic Environment
Cholera caused by toxigenic Vibrio cholerae is a major public health problem confronting developing countries, where outbreaks occur in a regular seasonal pattern and are particularly associated with poverty and poor sanitation. It is generally accepted that seven distinct pandemics of cholera have occurred since the onset of the first pandemic in 1817. Again Vibrio cholerae is capable of surviving in aquatic environments for extended periods and is considered as autochthonous species in estuarine and brackish waters. Therefore, the present study was designed to isolate V. cholerae from natural environmental samples subsequently identified by conventional and molecular biological techniques. A total number of 10 isolates were included randomly in this study based on their initial identification. The serotypes of the isolates were determined by serological test (slide agglutination) and the number of serotypes O1, O139 and non-O1/O139 were 3, 2 and 5 respectively which were reconfirmed by PCR method. Finally, the toxigenicity of the isolates was analyzed by multiplex PCR method and five (5) isolates were found to contain the ctx gene, the major virulence factor of V. cholerae. Key Words: Vibrio cholerae, Simplex PCR, Multiplex PCR, Serotypes, Toxigenicity.   doi:10.3329/sjps.v1i1.1811 S. J. Pharm. Sci. 1(1&2): 69-75
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