Multiplex PCR for Detection of Lactococcus garvieae, Streptococcus iniae and S. dysgalactiae in Cultured Yellowtail
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概要
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Streptococcosis is one of serious disease in cultured yellowtail, <I>Seriola quinqueradiata</I>, amberjack, <I>S. dumerili</I> and salmonids, and economically is problematic. <I>Streptococcus iniae</I>, is known as a pathogen of the marine and freshwater fishes. On the other hand, <I>Lactococcus garvieae</I>, and recently <I>S. dysgalactiae</I> are known as the pathogens of the yellowtail and amberjak. Mixed infection due to the pathogens, however, occurs very often in yellowtail. Therefore, the correct diagnosis is sometimes confused. In an attempt to elucidate the main pathogen, a multiplex PCR was newly designed in this study. The new multiplex PCR assays involves amplifying the three multiple gene products in a single reaction based on primers designed from the 16S rRNA, 16S-23S rDNA intergenic spacer region and lactate oxidase (<I>lctO</I>) genes of <I>L. garvieae, S. dysgalactiae</I> and <I>S. iniae</I>, respectively. The specificity of the multiplex PCR using the primer sets was confirmed bythe fact that specific bands were only amplified equivalent to 1, 100, 870 and 259-bp for <I>L. gayvieae</I>, <I>S. iniae</I>, and <I>S. dysgalactiae</I>, respectively. In addition, the specific positive amplifications in used all templates were consistently observed only for each corresponding pathogen. Multiplex PCR did not produce any non-specific amplification products when tested against pure DNAs, bacterial suspensions or tissue homogenates from rainbow trout, <I>Oncorhynchus mykiss</I>, artificially infected with the three pathogens. Multiplex PCR was an effective tool for the rapid and specific detection of <I>L. gravieae, S. dysgalactiae</I> and <I>S. iniae</I> from fish tissues.
- 日本水産増殖学会の論文
日本水産増殖学会 | 論文
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