鹅细小病毒LAMP快速检测方法的建立

摘要

By using PrimerExplorer V3 software, a series of specific primers were designed based on the nucleotide sequences of goose parvovirus (GPV) VP3 gene available in GenBank. These primers targeted 6 distinct conserved sequences of VP3 gene, and serve primers to perform a loop-mediated isothermal amplification (LAMP). These primers were used to establish rapid LAMP detection of GPV. The results showed the LAMP method specifieally amplified GPV but not other common avian disease DNA viruses;The method was 100 times more sensitive than the conventional PCR method. The amplification could be finished within 1.5 h with good specificity and high sensitivity. With the addition of SYBR GREEN dye in the reaction mixture, the results could be differentiated by naked eyes. The results also proved that LAMP method is a reliable and accurate assay for rapid detection of goose parvovirus. This assay offers a new technique and alternative for detecting goose parvovirus.%根据GenBank中登录的鹅细小病毒(GPV)VP3基因序列,利用PrimerExplorer V3软件在序列保守区域没计1套特异识别VP3基因序列中6个不同区段的环介导恒等温扩增(LAMP)引物,并以此套引物建立一种基于LAMP技术的GPV检测方法.结果表明,该方法灵敏度是普通PCR方法的100倍,只能特异性扩增GPV DNA,其引物对于其他常见禽DNA病毒均无特异性扩增.在反应体系中加入SYBR GREEN I染料后,可通过肉眼观察有无荧光直接判定结果.该方法特异性强、灵敏度高,重复性好,为快速检测鹅细小病毒提供了新方法和新思路.