Hoshino K, Yamasaki S, Mukhopadhyay A K, Chakraborty S, Basu A, Bhattacharya S K, Nair G B, Shimada T, Takeda Y
Research Institute, International Medical Center of Japan, Tokyo.
FEMS Immunol Med Microbiol. 1998 Mar;20(3):201-7. doi: 10.1111/j.1574-695X.1998.tb01128.x.
A multiplex polymerase chain reaction assay was developed for concurrent detection of rfb sequences specific for the O1 and the O139 serogroups of Vibrio cholerae and for ctxA specific sequences. The multiplex PCR assay was found to be highly specific and sensitive and was capable of detecting 65 cfu and 200 cfu per assay of V. cholerae O1 and O139, respectively. Evaluation of the multiplex PCR assay using 121 stool samples from patients admitted to the Infectious Diseases Hospital, Calcutta, showed the assay to be 100% sensitive and 95.2% specific when the culture method was taken as the standard. In addition to the 38 PCR positive stool samples, an additional four samples which were negative by culture method but positive by PCR assay belonged to the O139 serogroup. All the 38 stool samples positive for either O1 or O139 serogroup by PCR assay were also positive for the ctxA amplicon indicating that the O1 and O139 V. cholerae strains have the genetic potential of producing cholera toxin.
开发了一种多重聚合酶链反应检测方法，用于同时检测霍乱弧菌O1和O139血清群特异的rfb序列以及ctxA特异序列。发现该多重聚合酶链反应检测方法具有高度特异性和敏感性，每次检测分别能够检测到65个菌落形成单位（cfu）的霍乱弧菌O1和200个cfu的霍乱弧菌O139。使用来自加尔各答传染病医院收治患者的121份粪便样本对该多重聚合酶链反应检测方法进行评估，结果表明，以培养方法作为标准时，该检测方法的敏感性为100%，特异性为95.2%。除了38份聚合酶链反应阳性的粪便样本外，另外4份培养方法检测为阴性但聚合酶链反应检测为阳性的样本属于O139血清群。通过聚合酶链反应检测O1或O139血清群呈阳性的所有38份粪便样本，其ctxA扩增子也呈阳性，这表明霍乱弧菌O1和O139菌株具有产生霍乱毒素的遗传潜力。
