New Validated Rapid Screening Methods for Identifying Kudoa septempunctata in Olive Flounder (Paralichthys olivaceus)

Japanese Journal of Infectious Diseases

Yoshiko Sugita-Konishi⁴  Koh-ichiro Mori²  Makoto Kuroda¹  Takahiro Ohnishi⁵  Yutaka Fukuda³  Akiko Yamazaki⁵  Toyohiko Namba⁶  Toru Mekata² 

[1] Pathogen Genomics Center, National Institute of Infectious Diseases;National Research Institute of Aquaculture;Fisheries Research Division, Oita Prefectural Agriculture, Forestry and Fisheries Research Center;Graduate School of Environmental Health Science, Azabu University;Division of Microbiology, National Institute of Health Sciences;Incorporated Foundation Tokyo Kenbikyo-in

关键词: Kudoa septempunctata;    LAMP;    NASBA-nucleic acid chromatography;   

DOI  :  10.7883/yoken.JJID.2014.133

学科分类：传染病学

来源: National Institute of Infectious Diseases

【 摘 要 】

References(5)Cited-By(4)Kudoa septempunctata is a newly identified causative agent of foodborne diseases associated with consuming raw olive flounder. Qualitative PCR and quantitative real-time PCR have been used as notification methods to identify K. septempunctata in Japan. However, these methods require expensive equipment and are time-consuming (2–3 h for screening). To address these problems, in this study, we developed new rapid and simple methods using real-time loop-mediated isothermal amplification (LAMP) and nucleic acid sequence based amplification-nucleic acid chromatography (NASBA-NAC). Using these methods, the total procedure required approximately 45 min and did not require any expensive equipment. With regard to validating these new methods in comparison with the notification methods used in Japan, we performed an inter-laboratory study of 5 laboratories using samples that included olive flounders infected with 4 different amounts of K. septempunctata. These results demonstrated that the sensitivity of NASBA-NAC was equivalent to that of qualitative PCR, and that the sensitivity of real-time LAMP was equivalent to that of quantitative real-time PCR, which indicated that these new methods were acceptable screening methods for identifying K. septempunctata.

【 授权许可】

Unknown