Detection of urovirulence factors in Escherichia coli by multiplex polymerase chain reaction |
| |
Authors: | Shingo Yamamoto Akito Terai Kazuyo Yuri Hisao Kurazono Yoshifumi Takeda Osamu Yoshida |
| |
Affiliation: | Department of Urology, Faculty of Medicine, Kyoto University, Kawaharacho 54, Shogoin, Sakyo-ku, Kyoto 606-01, Japan;Department of Microbiology, Faculty of Medicine, Kyoto University, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-01, Japan;Dainippon Pharmaceutical Co., Ltd., 6-8 Doshomachi, 2-chome, Chuo-ku, Osaka 541, Japan;Research Institute, International Medical Center of Japan, 1-21-1, Toyama, Shinjuku-ku, Tokyo 162, Japan |
| |
Abstract: | Abstract 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 × 103 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 . |
| |
Keywords: | Escherichia coli Urovirulence factor Polymerase chain reaction (PCR) |
|
|