Japanese Journal of Infectious Diseases
Online ISSN : 1884-2836
Print ISSN : 1344-6304
ISSN-L : 1344-6304
New Validated Rapid Screening Methods for Identifying Kudoa septempunctata in Olive Flounder (Paralichthys olivaceus)
Yoshiko Sugita-KonishiYutaka FukudaKoh-ichiro MoriToru MekataToyohiko NambaMakoto KurodaAkiko YamazakiTakahiro Ohnishi
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JOURNALS FREE ACCESS Advance online publication

Article ID: JJID.2014.133

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Abstract

Kudoa septempunctata is a new causative agent of foodborne diseases associated with the consumption of raw olive flounder. Qualitative PCR and quantitative real-time PCR have been developed as notification methods to detect K. septempunctata in Japan; however, these methods require expensive equipment and are time-consuming, taking two to three hours. In order to address these problems, in this study, we developed new rapid and simple methods using real-time LAMP and NASBA-nucleic acid chromatography (NASBA-NAC). In these methods, the total procedure requires approximately 45 minutes, without the need for any expensive equipment. With respect to the validation of these new methods in comparison to the notification methods used in Japan, an inter-laboratory study was performed among five laboratories using samples that included olive flounders infected with four different amounts of K. septempunctata. The results demonstrated that the sensitivity of NASBA-NAC was equivalent to that of qualitative PCR, while the sensitivity of real-time LAMP was equivalent to that of quantitative real-time PCR, suggesting that these new methods are acceptable screening methods for identifying K. septempunctata.

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