Open AccessDetection of urovirulence factors in Escherichia coli by multiplex polymerase chain reaction
Author(s) -
Yamamoto Shingo,
Terai Akito,
Yuri Kazuyo,
Kurazono Hisao,
Takeda Yoshifumi,
Yoshida Osamu
Publication year - 1995
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.1995.tb00179.x
Subject(s) - multiplex polymerase chain reaction , escherichia coli , biology , polymerase chain reaction , virulence , microbiology and biotechnology , aerobactin , bacterial adhesin , hemolysin , fimbria , virulence factor , multiplex , gene , enterobacteriaceae , genetics
Primers to amplify the genes encoding the virulence factors of uropathogenic Escherichia coli , such as pilus associated with pyelonephritis ( pap ), haemolysin ( hly ), aerobactin ( aer ) and cytotoxic necrotizing factor 1 ( cnf 1) genes, were designed. The above primers along with previously reported primers for S fimbriae ( sfa ) and afimbrial adhesin I ( afaI ) genes were combined to develop a multiplex polymerase chain reaction (PCR) for detection of the respective virulence factors and for the identification of uropathogenic E. coli . The multiplex PCR to detect pap, sfa, afa I, hly, aer and cnf 1 genes was highly specific and the sensitivity was found to be about 5 × 10 3 colony forming units of E. coli per ml. A total of 194 E. coli strains isolated from patients with simple acute cystitis were examined by the multiplex PCR and the results were in complete agreement with that obtained by DNA colony hybridization test. The multiplex PCR developed was, therefore, concluded to be a useful, sensitive and rapid assay system to identify uropathogenic E. coli .