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Highly specific and rapid detection of Hamiltonella defense, a secondary endosymbiont of Bemisia tabaci, which plays critical roles on TYLCV transmission by loop-mediated isothermal amplification (LAMP) assay

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  • URLhttps://db.koreascholar.com/Article/Detail/317795
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한국응용곤충학회 (Korean Society Of Applied Entomology)
초록

Loop-mediated isothermal amplification (LAMP) is a rapid, specific, cost-effective detection method by amplifying nucleic acid under isothermal conditions. In this study, we used LAMP for detection of Hamiltonella defensa that lives as a facultive endosymbiont of whitefly ‘Bemisia tabaci’. We designed the Hamiltonella-specific primers by targeting 16S ribosomal RNA gene and validated the specificity of one primer set. To find the optimum temperature for our primer set, the LAMP reaction was held at the temperature, 60℃, 62℃ and 65℃. As a result, 62℃ was the optimum reation temperature for LAMP reaction. Specificity of primer set was tested by the reaction to both Trialeurodes vaporariorum and B. tabaci. After the whole procedure, the amplicons by LAMP were visualized by adding SYBR Green to the reaction tube.

저자
  • Eun-Yong Choi(Department of Genetic Engineering, Sungkyunkwan University)
  • Haneul Seo(Department of Genetic Engineering, Sungkyunkwan University)
  • Eui-Joon Kil(Department of Genetic Engineering, Sungkyunkwan University)
  • Hee-Seong Byun(Department of Genetic Engineering, Sungkyunkwan University)
  • Jae-Kyoung Shim(Institute of Plant Medicine, Kyungpook National University)
  • Ji-Kwang Kim(Research and Development Bureau, Chungcheongnam-do Agricultural Research and Extension Services)
  • Jung-Hwan Lee(Institute for Bioresources Research, Gyeongsangbuk-do Agricultural Research and Extension Services)
  • Gwan-Seok Lee(Crop Protection Division, National Institute of Agricultural Science, Rural Development Administration)
  • Chang-Seok Kim(Crop Protection Division, National Institute of Agricultural Science, Rural Development Administration)
  • Hong-Soo Choi(Crop Protection Division, National Institute of Agricultural Science, Rural Development Administration)
  • Sukchan Lee(Department of Genetic Engineering, Sungkyunkwan University)
  • Kyeong-Yeoll Lee(Institute of Plant Medicine, Kyungpook National University)