Parasites, Hosts and Diseases

  • 제52권3호
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  • Pages.305-310
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  • 2014
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  • 2982-5164(pISSN)
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  • 1738-0006(eISSN)
대한기생충학·열대의학회 (The Korea Society for Parasitology and Tropical Medicine)
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Development of Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Azumiobodo hoyamushi (Kinetoplastea)

  • Song, Su-Min (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Sylvatrie-Danne, Dinzouna-Boutamba (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Joo, So-Young (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Shin, Yun Kyung (Aquaculture Management Division, National Fisheries Research & Development Institute) ;
  • Yu, Hak Sun (Department of Parasitology, School of Medicine, Pusan National University) ;
  • Lee, Yong-Seok (Department of Life Science and Biotechnology, College of Natural Sciences, Soonchunhyang University) ;
  • Jung, Ji-Eon (Department of Life Science and Biotechnology, College of Natural Sciences, Soonchunhyang University) ;
  • Inoue, Noboru (National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine) ;
  • Lee, Won Kee (Department of Preventive Medicine, Kyungpook National University School of Medicine) ;
  • Goo, Youn-Kyoung (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Chung, Dong-Il (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine) ;
  • Hong, Yeonchul (Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine)
  • 투고 : 2014.02.28
  • 심사 : 2014.04.04
  • 발행 : 2014.06.30
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초록

Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.

키워드

참고문헌

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피인용 문헌

  1. Measurement of Tunic Hardness in an Edible Ascidian, Halocynthia roretzi, with Remarks on Soft Tunic Syndrome vol.35, pp.6, 2014, https://doi.org/10.2108/zs180072