Multiplex Multilocus Variable-Number Tandem-Repeat Analysis for Typing of Pandemic Vibrio parahaemolyticus O1:KUT Isolates.

Pandemic O3:K6 Vibrio parahaemolyticus emerged in 1996. Since then, this strain of pathogen and its serovariants (predominantly O1:KUT [untypable], O1:K25 and O4:K68) have caused gastroenteritis worldwide. Owing to the limitation in established K antisera, tracking the sources of KUT for epidemiological investigation is difficult. Therefore, the effective molecular typing is required to discriminate the strains. The aim of this study was to develop a multiplex multilocus variable-number tandem-repeat analysis (MLVA) assay for typing pandemic V. parahaemolyticus, including various O1:KUT isolates. The assay was based on the analysis of four variable number tandem repeat loci. Forty-six pandemic isolates, including O1:KUT, O1:K25, and O3:K6, were investigated. MLVA generated 38 distinct MLVA profiles, whereas only 16 types were obtained from pulsed-field gel electrophoresis (PFGE). In this work, MLVA resolved the 12 isolates of O1:KUT obtained in 2001-2005 with identical PFGE patterns into unique profiles. Our data indicated that multiplex MLVA developed in this study has high discriminatory power (D = 0.99), and is superior to PFGE for distinct pandemic V. parahaemolyticus, including O1:KUT isolates.