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Improved One-step Colony PCR Detection of Vibrio harveyi

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Title: Improved One-step Colony PCR Detection of Vibrio harveyi
Authors: Fukui, Youhei Browse this author
Sawabe, Tomoo Browse this author →KAKEN DB
Keywords: Vibrio harveyi
16S rRNA gene
colony PCR
Issue Date: Mar-2007
Publisher: Japanese Society of Microbial Ecology
Journal Title: Microbes and Environments
Volume: 22
Issue: 1
Start Page: 1
End Page: 10
Publisher DOI: 10.1264/jsme2.22.1
Abstract: Vibrio harveyi strains are pathogenic to a wide range of marine fish and shellfish, having a significant negative economic impact on aquaculture worldwide. However, a reliable and rapid method of detecting V. harveyi has yet to be established. We aimed to construct an improved one-step detection method for V. harveyi. Reanalysis of 16S rRNA gene sequences of type strains of V. harveyi revealed a unique consensus region compared to related Vibrio species. Using a VHARF-VHARR primer set from this region, a V. harveyi-specific PCR-based detection method was established and could differentiate V. harveyi from related species such as V. campbellii, V. rotiferianus, and V. alginolyticus. Furthermore, the new method (optimal amplification with 20 core cycles of (94℃-30 s, 60℃-30 s, 72℃-30 s)) could be applied to the identification of V. harveyi strains of colonies. This PCR was able to detect V. harveyi grown on plates in the environment within 3 days without bacterial isolation, DNA extraction, or the assistance of biochemical tests. The specificity and rapidity of the detection is reliable enough to understand the ecology of V. harveyi in environments, and to predict outbreaks of mass mortality caused by V. harveyi in aquaculture.
Rights: 日本微生物生態学会 (Japanese Society of Microbial Ecology)
Type: article
Appears in Collections:水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 澤辺 智雄

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