Development of quantitative method by loop-mediated isothermal amplification (LAMP) using cycling probe.

2015 Fiscal Year Final Research Report

• Project/Area Number: 25460705
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Fujita Health University
• Principal Investigator: IHIRA Masaru 藤田保健衛生大学, 保健学研究科, 教授 (10290165)
• Co-Investigator(Kenkyū-buntansha): SUGIYAMA Hiroko 藤田保健衛生大学, 医学研究科, 研究員 (10387714) ; ENOMOTO Yoshihiko 藤田保健衛生大学, 医学研究科, 研究員 (00387713) ; YOSHIKAWA Tetsushi 藤田保健衛生大学, 医学部, 教授 (80288472)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: HHV-6A / HHV-6B / LAMP / cycling probe

Outline of Final Research Achievements

Aim of this study was to develop quantitative LAMP method for measurement of target DNA load by using cycling probe. We combine LAMP and cycling probe to develop new quantitative method. The cycling probe PCR had a broad, linear dynamic range of detection between 10 and 1000000 copies of viral DNA. Strong correlations were demonstrated between cycling probe PCR and Taqman real-time PCR. The optimized condition for cycling probe LAMP was confirmed using variable concentration of pH and probe. The cycling probe LAMP assay had excellent linear dynamic range between 1000 and 1000000 copies of viral DNA. However, cycling probe LAMP was less sensitive than Taqman real-time PCR. Result of detection from HHV6 DNA in serum by our developed dry reagent for LAMP were correspond with that of liquid reagent. Our cycling probe LAMP require improved sensitivity. However, combine dry reagent-LAMP and cycling probe suggest the possibility of rapid and simple DNA quantitation method.

Free Research Field

臨床ウイルス学