| HWANG Jinik,PARK So Yun,SUH Sung-Suk,PARK Mirye,LEE Sukchan,LEE Taek-Kyun. 2016. Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP). Acta Oceanologica Sinica, 35(8):44－50 |
| Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP) |
| Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP) |
| Received:August 11, 2015 Revised:December 16, 2015 |
| DOI：10.1007/s13131-016-0889-7 |
| Key words:viral hemorrhagic septicaemia virus (VHSV) marine birnavirus (MABV) polymerase chain reaction loop-mediated isothermal amplification |
| 中文关键词: |
| 基金项目:The grants from the Korea Institute of Ocean Science and Technology under contract No. PE99315 |
| Author Name | Affiliation | E-mail | | HWANG Jinik | South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea
Korea University of Science and Technology, Daejeon 34113, Republic of Korea | | | PARK So Yun | South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea | | | SUH Sung-Suk | South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea | | | PARK Mirye | South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea
Korea University of Science and Technology, Daejeon 34113, Republic of Korea | | | LEE Sukchan | Department of Genetic Engineering, Sungkyunkwan University, Suwon 16419, Republic of Korea | | | LEE Taek-Kyun | South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea
Korea University of Science and Technology, Daejeon 34113, Republic of Korea | tklee@kiost.ac |
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| Abstract: |
| Viral hemorrhagic septicemia virus (VHSV) and marine birnavirus (MABV) are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogen infection. The loop-mediated isothermal amplification (LAMP) of DNA is currently one of the most commonly used molecular diagnostic tools, as it is simple, quick, and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fish was developed. Using a set of synthesized primers matching a specific region of the genome, the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficient detection of VHSV and MABV infection in the wild fish, Paralichthys olivaceus. Our results demonstrate the simplicity and convenience of this method for the detection of viral infection in aquatic organisms. |
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