Multi-Detection by Target Mixed Loop-Mediated Isothermal Amplification
Kentaro Nagamine *
Hiroshima International University, 5-1-1 Hirokoshingai, kure, Hiroshima 737-0112, Japan
Yoko Kuzuhara
Eiken Chemical Co., Ltd., 1381-3 Shimoishigami, Ohtawara, Tochigi 324-0036, Japan
Tsugunori Notomi
Eiken Chemical Co., Ltd., 1381-3 Shimoishigami, Ohtawara, Tochigi 324-0036, Japan
Junichi Takino
Hiroshima International University, 5-1-1 Hirokoshingai, kure, Hiroshima 737-0112, Japan
Takamitsu Hori
Hiroshima International University, 5-1-1 Hirokoshingai, kure, Hiroshima 737-0112, Japan
Hidetoshi Kanda
Eiken Chemical Co., Ltd., 1381-3 Shimoishigami, Ohtawara, Tochigi 324-0036, Japan
*Author to whom correspondence should be addressed.
Abstract
Loop-mediated isothermal amplification (LAMP), a nucleic acid amplification technology, has been characterized as having a short reaction time while generating a large amount of DNA product under isothermal conditions. Here, we found that LAMP is able to amplify more than one target simultaneously by using a mixture of human immunodeficiency virus-1, hepatitis B virus, and hepatitis C virus primers, a process we have named target mixed LAMP. Target mixed LAMP and detection can be performed in less than 1 h. In this study, we also used target mixed LAMP to successfully amplify GAPDH, gamma-actin, TATA box binding protein, and haptoglobin from pooled cDNA derived from mouse tissues. Furthermore, using eight plasmid vectors as templates, target mixed LAMP is able to amplify all targets simultaneously. These findings reveal the usefulness of LAMP for the diagnosis of infectious disease as well as for conducting gene expression analysis.
Keywords: LAMP, infectious disease, virus, diagnosis, magnesium pyrophosphate, DNA amplification