Loop-mediated isothermal amplification for rapid detection of acute viral necrobiotic virus in scallop Chlamys farreri

Acta Virologica
W C RenY Y Cai

Abstract

Loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive detection of Acute viral necrobiotic virus (AVNV) in scallop Chlamys farreri was developed and evaluated. Four primers recognizing six targets on distinct AVNV DNA sequences were designed and the LAMP reaction was carried out in a water bath. Reaction temperature and time were optimized at 64 degrees C for 60 mins and LAMP products were detected using agarose gel electrophoresis and visual assessment. Confirmation of the expected LAMP products was performed with MboI restriction enzyme analysis. The detection limit of LAMP assay was as low as 1 fg AVNV DNA and accordingly, this assay was 100 times more sensitive than conventional PCR technique. A comparative evaluation of 20 samples using the LAMP and PCR assays revealed a complete accord in positivity or negativity for AVNV. These results indicate that the LAMP assay is simple, sensitive, specific, and has a great potential for detection of AVNV in the laboratory and field.

Citations

Jun 5, 2014·Applied Biochemistry and Biotechnology·Mohammad Amin AlmasiMehdi Aghapour-ojaghkandi
May 29, 2012·Journal of Virological Methods·Jing-Zhe JiangJiang-Yong Wang
May 23, 2012·Journal of Virological Methods·Jing-Zhe JiangJiang-Yong Wang
Mar 31, 2015·Applied Biochemistry and Biotechnology·Mohammad Amin AlmasiSeyed Mohammad Hosseyni-Dehabadi
Apr 29, 2020·Micromachines·Alana Torres VidalHieu Bui
Feb 20, 2021·Materials Science for Energy Technologies·Victoria Xin Ting ZhaoXiaodong Zhou

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