Environmental investigation of potentially pathogenic Vibrio parahaemolyticus in the Seto-Inland Sea, Japan.
Environmental investigation of potentially pathogenic Vibrio parahaemolyticus in the Seto-Inland Sea, Japan.
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DOI：
10.1111/j.1574-6968.2002.tb11064.x
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发表时间：
2002-02
影响因子：
2.1
通讯作者：
M. J. Alam;K. Tomochika;S. Miyoshi;S. Shinoda
中科院分区：
文献类型：
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作者：
M. J. Alam;K. Tomochika;S. Miyoshi;S. Shinoda
在寒冷的天气季节从日本滨海采集海水和有机物(沉没在海水中的任何底物上的活的和/或死亡的物质)，并用从样品的增菌培养中分离的DNA进行分析，以确定副溶血性弧菌的密度和病原菌的出现情况，通过聚合酶链式反应(PCR)将病原菌定义为具有tdh和/或trh基因的菌株。用种特异性毒素R引物的最可能数(MPN)-PCR技术检测副溶血性弧菌(每100毫升水或10克有机物质的细胞密度为3~240个/100ml水或10g有机物质)的阳性率约为95%，而用常规的MPN-培养技术(每100毫升水或10g有机物质的细胞密度为3~240个/100毫升或10g有机物质)的阳性率仅为40%。此外，用MPN-PCR技术检测，tdh和trh基因的阳性率分别为55%和20%。常规MPN培养法未分离到tdh和trh基因阳性菌株。MPN培养和MPN-PCR技术之间的检测差异似乎是显著的，这可能是由于不同的检测灵敏度和其他因素所致。
Seawater and organic material (live and/or dead matter deposited on any substratum submersed in seawater) were collected during the cool weather season from a coast of the Seto-Inland Sea, Japan, and analyzed to determine Vibrio parahaemolyticus densities and the occurrence of pathogenic strains, defined as those possessing tdh and/or trh genes by the polymerase chain reaction (PCR), using isolated DNA from enrichment culture of the samples. About 95% of the samples were positive for V. parahaemolyticus (with densities of 3 to >1400 cells per 100 ml water or 10 g organic samples) by the most-probable-number (MPN)-PCR technique with species-specific toxR primers, but only 40% were positive by the conventional MPN-culture technique (with densities ranging from 3 to 240 cells per 100 ml water or 10 g organics). Furthermore, the tdh and trh genes were positive in 55% and 20% of samples, respectively, by the MPN-PCR technique. No tdh and trh gene-positive strains were isolated by the conventional MPN-culture procedure. The difference in detection between the MPN-culture and the MPN-PCR techniques appeared to be significant and may be attributed to different detection sensitivities and other factors.