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Rapid Detection of the pathogenic agent of Bacterial white enteritis of Larval and Juvenile Stages in Olive flounder (Paralichthys olivaceus)  

Mun, Yeong-Geon (제주대학교)
Park, Geun-Tae (제주대학교)
Son, Hong-Ju (밀양대학교)
Lee, Sang-Hyeon (신라대학교)
Lee, Jeong-Min (남부대학교)
Heo, Mun-Su (제주대학교)
Publication Information
Journal of fish pathology / v.17, no.3, 2004 , pp. 159-169 More about this Journal
Abstract
Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.
Keywords
Vibrio ichthyoenteri, bacterial white enteritis, 16S-23S rRNA ISR, tRNA gene, DNA sequencing
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