| Jun Young Kim | 2 Articles |
Objectives
A multiplex real-time polymerase chain reaction (RT-PCR) method was developed for the identification of three <i>Vibrio</i> species: <i>Vibrio cholerae</i>, <i>Vibrio parahaemolyticus</i>, and <i>Vibrio vulnificus</i>. Methods Specific primers and probes targeting the <i>hlyA</i>, <i>tlh</i>, and <i>vvhA</i> genes were selected and used for multiplex real-time PCR to confirm the identification of <i>V. cholerae</i>, <i>V. parahaemolyticus</i>, and <i>V. vulnificus</i>, respectively. This method was applied to screen <i>Vibrio</i> species from environmental samples and combining it with a culture-based method, its effectiveness was evaluated in comparison with culture-based methods alone. Results Specific PCR fragments were obtained from isolates belonging to the target species, indicating a high specificity of this multiplex real-time PCR. No cross-reactivity with the assay was observed between the tested bacteria. The sensitivity of the multiplex real-time PCR was found to have a lower limit of 10<sup>4</sup> colony-forming units/reaction for all three <i>Vibrio</i> species. The combination strategy raised the isolation ratio of all three <i>Vibrio</i> species 1.26- to 2.75-fold. Conclusion This assay provides a rapid, sensitive, and specific technique to detect these three <i>Vibrio</i> species in the environment. Citations
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Objectives
<i>Vibrio vunificus</i> is known to cause septicemia and severe wound infections in patients with chronic liver diseases or an immuno-compromised condition. We carried out the molecular characterization of <i>V. vulnificus</i> isolates from human <i>Vibrio</i> septicemia cases based on pulsed-field gel electrophoresis (PFGE) using NotI and SfiI. Methods and Results PFGE was used to characterize a total of 78 strains from clinical cases after NotI or SfiI digestion. The geographical distribution of PFGE patterns for the strains from the southern part of Korea, a high-risk region for <i>Vibrio</i> septicemia, indicated that the isolates from southeastern Korea showed a comparatively higher degree of homology than those from southwestern Korea. Conclusions We report the genetic distribution of <i>V. vulnficus</i> isolated from <i>Vibrio</i> septicemia cases during 2000–2004 in Korea. This method has potential use as a subspecies-typing tool for <i>V. vulnificus</i> strains isolated from distant geographic regions. Citations
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