MOLECULAR EPIDEMIOLOGIC EVIDENCE FOR ASSOCIATION OF THERMOSTABLE DIRECT HEMOLYSIN (TDH) AND TDH-RELATED HEMOLYSIN OF VIBRIO-PARAHAEMOLYTICUS WITH GASTROENTERITIS
MOLECULAR EPIDEMIOLOGIC EVIDENCE FOR ASSOCIATION OF THERMOSTABLE DIRECT HEMOLYSIN (TDH) AND TDH-RELATED HEMOLYSIN OF VIBRIO-PARAHAEMOLYTICUS WITH GASTROENTERITIS
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DOI：
10.1128/iai.58.11.3568-3573.1990
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发表时间：
1990-11-01
影响因子：
3.1
通讯作者：
NISHIBUCHI, M
中科院分区：
文献类型：
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作者：
SHIRAI, H;ITO, H;NISHIBUCHI, M
副溶血性弧菌的热稳定直接溶血素（TDH）引起的神奈川现象几乎只与临床菌株有关，TDH被认为是一种重要的毒力因子。然而，从腹泻患者中分离的神奈川现象阴性菌株最近显示产生TDH相关溶血素（TRH）。我们用杂交分析的方法研究了编码TDH的tdh基因和编码TRH的trh基因在弧菌中的分布。用tdh基因特异性探针和一种新构建的trh基因特异性探针，对285株副溶血性弧菌进行了tdh基因和trh基因的DNA菌落杂交检测。为了评估TRH的重要性，纳入了许多神奈川现象阴性的临床菌株（占所有菌株的35.4%）。在214株副溶血性弧菌临床分离株中，112株（52.3%）仅携带tdh基因，52株（24.3%）仅携带trh基因，24株（11.2%）同时携带tdh和trh基因。Southern印迹杂交分析证实了tdh和trh基因在这24株菌中的共存。在71株环境菌株中，有5株（7.0%）与trh基因探针呈弱杂交，未与tdh基因探针杂交。这些结果表明，TRH和TDH是副溶血性弧菌的重要毒力因子。在118株其他弧菌中检测了trh基因，只有1株Vibrioissii产生了非常弱的杂交信号。在48株具有代表性的trh基因阳性的副溶血性弧菌中，用敏感的酶联免疫吸附试验检测，只有18株（37.5%）菌株在培养基中产生TRH。
The Kanagawa phenomenon induced by the thermostable direct hemolysin (TDH) of Vibrio parahaemolyticus is almost exclusively associated with clinical strains, and TDH has been considered an important virulence factor. However, Kanagawa phenomenon-negative strains isolated from patients with diarrhea have recently been shown to produce TDH-related hemolysin (TRH). We studied the distribution of the tdh gene encoding TDH and the trh gene encoding TRH in vibrios by hybridization analyses. The presence or absence of the tdh gene and the trh gene in 285 strains of V. parahaemolyticus was examined by the DNA colony hybridization test with a tdh gene-specific probe and a newly constructed trh gene-specific probe. For assessment of the importance of TRH, many Kanagawa phenomenon-negative clinical strains (35.4% of all strains) were included. Of 214 clinical strains of V. parahaemolyticus, 112 strains (52.3%) had the tdh gene only, 52 strains (24.3%) had the trh gene only, and 24 strains (11.2%) carried both the tdh and the trh gene. The coexistence of the tdh and trh genes in these 24 strains was confirmed by Southern blot hybridization analysis. Of 71 environmental strains, 5 strains (7.0%) hybridized very weakly with the trh gene probe and none hybridized with the tdh gene probe. These results suggest that TRH as well as TDH is an important virulence factor of V. parahaemolyticus. Among 118 strains of other Vibrio species examined for the trh gene, only 1 strain of Vibrio furnissii gave a very weak hybridization signal. Among 48 representative trh gene-positive strains of V. parahaemolyticus, only 18 strains (37.5%) were found to produce TRH in culture medium when examined by a sensitive enzyme-linked immunosorbent assay method.