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Diversity of DNA sequences among Vibrio cholerae O139 Bengal detected by PCR-based DNA fingerprinting
Authors:Sou-ichi Makino  Takayuki Kurazono  Yusuke Okuyama  Toshio Shimada  Yumiko Okada  Chihiro Sasakawa
Institution:Department of Veterinary Public Health, The Institute of Public Health, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108, Japan; Saitama Institute of Public Health, Saitama 338, Japan; The National Institute of Health, Tokyo 108, Japan; Department of Bacteriology, Institute of Medical Science, University of Tokyo, Tokyo 108, Japan
Abstract:Abstract Vibrio cholerae O139, a causative agent of a large epidemic of cholera-like illness, has suddenly emerged and spread widely over several months. To investigate the characteristics unique to O139, traditional typing techniques for V. cholerae , such as biochemical characteristics, antibiotic susceptibility and detection of toxin production, were performed, with the result that 145 O139 strains, except for two O139 strains isolated from Argentina and Germany, were indistinguishable from O1 strains. Thus, in order to clarify the genetical relatedness among O139 strains, and between O139 and O1 strains, the RAPD (random amplified polymorphic DNA) DNA fingerprinting method was undertaken. Although the RAPD arrays in five O139 isolates from Vellore with one arbitrary primer were slightly different from the other O139 strains, the RAPD patterns of the 145 forty-five O139 strains except for two O139 strains from Argentina and Germany were quite similar to each other, but were different from those of O1 strains, indicating that those O139 epidemic strains are closely related to each other regardless of their place of isolation. Furthermore, the RAPD patterns of the O139 strains resembled those of E1 Tor strains rather than classical strain, and a small change in the RAPD pattern of O139 strains occurred during subculture for 200 generations. These results taken together suggested that O139 V. cholerae have emerged from a common origin associated with the E1 Tor strain.
Keywords:Polymerase chain reaction  Random amplified polymorphic DNA  Arbitrary primer  Fingerprinting              Vibrio cholerae
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