Efficient detection of pathogen virus in sand dabs, <i>Paralichthys olivaceus</i> using loop-mediated isothermal amplification (LAMP)

HWANG Jinik; PARK So Yun; SUH Sung-Suk; PARK Mirye; LEE Sukchan; LEE Taek-Kyun

doi:10.1007/s13131-016-0889-7

Volume 35 Issue 8

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Article Navigation > Acta Oceanologica Sinica > 2016 > 35(8): 44-50

HWANG Jinik, PARK So Yun, SUH Sung-Suk, PARK Mirye, LEE Sukchan, LEE Taek-Kyun. Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP)[J]. Acta Oceanologica Sinica, 2016, 35(8): 44-50. doi: 10.1007/s13131-016-0889-7

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HWANG Jinik, PARK So Yun, SUH Sung-Suk, PARK Mirye, LEE Sukchan, LEE Taek-Kyun. Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP)[J]. Acta Oceanologica Sinica, 2016, 35(8): 44-50. doi: 10.1007/s13131-016-0889-7

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Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP)

doi: 10.1007/s13131-016-0889-7

1.
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
2.
South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea
3.
Department of Genetic Engineering, Sungkyunkwan University, Suwon 16419, Republic of Korea

Received Date: 2015-08-11
Rev Recd Date: 2015-12-16

Abstract

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.
- viral hemorrhagic septicaemia virus (VHSV),
- marine birnavirus (MABV),
- polymerase chain reaction,
- loop-mediated isothermal amplification

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