Hara-Kudo Yukiko, Sugiyama Kanji, Nishibuchi Mitsuaki, Chowdhury Ashrafuzzaman, Yatsuyanagi Jun, Ohtomo Yoshimitsu, Saito Akinobu, Nagano Hidetoshi, Nishina Tokuhiro, Nakagawa Hiroshi, Konuma Hirotaka, Miyahara Michiko, Kumagai Susumu
Department of Biomedical Food Research, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan.
Appl Environ Microbiol. 2003 Jul;69(7):3883-91. doi: 10.1128/AEM.69.7.3883-3891.2003.
Although thermostable direct hemolysin (TDH)-producing Vibrio parahaemolyticus has caused many infections in Asian countries, the United States, and other countries, it has been difficult to detect the same pathogen in seafoods and other environmental samples. In this study, we detected and enumerated tdh gene-positive V. parahaemolyticus in Japanese seafoods with a tdh-specific PCR method, a chromogenic agar medium, and a most-probable-number method. The tdh gene was detected in 33 of 329 seafood samples (10.0%). The number of tdh-positive V. parahaemolyticus ranged from <3 to 93/10 g. The incidence of tdh-positive V. parahaemolyticus tended to be high in samples contaminated with relatively high levels of total V. parahaemolyticus. TDH-producing strains of V. parahaemolyticus were isolated from 11 of 33 tdh-positive samples (short-necked clam, hen clam, and rock oyster). TDH-producing strains of V. parahaemolyticus were also isolated from the sediments of rivers near the coast in Japan. Representative strains of the seafood and sediment isolates were examined for the O:K serovar and by the PCR method specific to the pandemic clone and arbitrarily primed PCR and pulsed-field gel electrophoresis techniques. The results indicated that most O3:K6 tdh-positive strains belonged to the pandemic O3:K6 clone and suggested that serovariation took place in the Japanese environment.
尽管产耐热直接溶血素(TDH)的副溶血性弧菌已在亚洲国家、美国及其他国家引发了诸多感染,但在海产品及其他环境样本中检测到同一病原体却并非易事。在本研究中,我们运用tdh特异性PCR方法、显色琼脂培养基及最大可能数法,对日本海产品中产tdh基因的副溶血性弧菌进行了检测与计数。在329份海产品样本中,有33份(10.0%)检测到tdh基因。产tdh的副溶血性弧菌数量在每10克<3至93株之间。在副溶血性弧菌总污染水平相对较高的样本中,产tdh的副溶血性弧菌的发生率往往较高。从33份tdh阳性样本中的11份(短颈蛤、文蛤和生蚝)分离出产TDH的副溶血性弧菌菌株。在日本沿海附近河流的沉积物中也分离出产TDH的副溶血性弧菌菌株。对海产品和沉积物分离株的代表性菌株进行了O:K血清型检测,并采用针对大流行克隆的PCR方法、任意引物PCR及脉冲场凝胶电泳技术进行检测。结果表明,大多数O3:K6 tdh阳性菌株属于大流行的O3:K6克隆,提示在日本环境中发生了血清型变异。