Establishment and Application of LAMP Detection Method of Bovine Viral Diarrhea Virus |
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Authors: | LI Jia-wei GUO Li YANG Yong WANG Jian-ke ZHANG Shu-qin ZHANG Jia-li CHENG Shi-peng |
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Institution: | 1. Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;2. Graduate School, Jilin Agricultural University, Changchun 130118, China |
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Abstract: | Since the 5'UTR gene (GenBank No.:AY278459.1) had 4 isolated regions,we designed a set of 4 LAMP primers to specifically recognize target gene sequences.This study developed a loop-mediated isothermal amplification (LAMP) method for detecting BVDV,using the pyrophosphate magnesium white precipitate for Real-time detection in LAMP reaction process of turbidity instrument,Real-time monitor liquid turbidity to determine result.The whole reaction lasted only 50 minutes at a constructed temperature of 63 ℃ to evaluate specificity,sensibility and repeatability of the method.The result demonstrated that the LAMP assay could only react with BVDV,its specificity was high;It could detect at least 10-6-fold diluted samples,which was 100 more sensitive than PCR assay;And repeatability was good.The simple,rapid,high siensitivity and specificity LAMP assay was a potential tool for the detection of BVDV in field conditions. |
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Keywords: | bovine viral diarrhea virus (BVDV) loop-mediated isothermal amplification method (LAMP) PCR |
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