Vicente A C, Coelho A M, Salles C A
Department of Genetics, Institute Oswaldo Cruz, Federal University of Rio de Janeiro, Brazil.
J Med Microbiol. 1997 May;46(5):398-402. doi: 10.1099/00222615-46-5-398.
Previously the heat-stable enterotoxin in Vibrio cholerae and V. mimicus has been detected by suckling mouse assay, a non-specific approach, and by DNA probes, a time-consuming method. This report describes a polymerase chain reaction (PCR) procedure for the detection of the stn (NAG-ST) and sto (O1-ST) gene sequences that is rapid and specific, allowing toxin gene molecular characterisation. A total of 34 V. cholerae and V. mimicus isolates was examined for ST and CT genes. The NAG-ST gene sequence was amplified in 13 of 22 non-O1/non-O139 V. cholerae and three of five V. mimicus strains. A new enterotoxin gene sequence pattern was found with MseI and TaqI restriction endonuclease PCR fragment digestion of two V. cholerae isolates, in addition to the pattern anticipated from the Genbank sequence, and found with the other ST+. These results show that ST-PCR detection is useful for the characterisation of V. cholerae and V. mimicus.
以前,霍乱弧菌和拟态弧菌中的热稳定肠毒素是通过乳鼠试验(一种非特异性方法)和DNA探针(一种耗时的方法)检测到的。本报告描述了一种用于检测stn(NAG-ST)和sto(O1-ST)基因序列的聚合酶链反应(PCR)程序,该程序快速且特异,可实现毒素基因的分子特征分析。总共对34株霍乱弧菌和拟态弧菌分离株进行了ST和CT基因检测。在22株非O1/非O139霍乱弧菌中的13株以及5株拟态弧菌中的3株中扩增出了NAG-ST基因序列。除了从Genbank序列预期的模式以及在其他ST+菌株中发现的模式外,通过对两株霍乱弧菌分离株的MseI和TaqI限制性内切酶PCR片段消化发现了一种新的肠毒素基因序列模式。这些结果表明,ST-PCR检测对于霍乱弧菌和拟态弧菌的特征分析很有用。