农业生物技术学报2011，Vol.19Issue(1)：186-190,5.DOI:10.3969/j.issn.1674-7968.2011.01.027
环介导恒温扩增技术快速检测对虾白斑综合征病毒方法的建立
Establishment of Loop-mediated Isothermal Amplification for Detection of White spot syndrome virus
摘要
Abstract
Loop-mediated isothermal amplification (LAMP) method is a novel gene detection technique with specificity, sensitive and rapidity, the products of which can be electrophoresed in 1.5％ agarose gels, or visually inspected by adding nucleic acid dye (SYBR Green Ⅰ ). In this paper, a one step loop-mediated isothermal amplification (LAMP) assay was developed for detection of White spot syndrome virus (WSSV). A set of primers were designed from the VP26 gene sequence of WSSV. The assay was established to amplify WSSV genomic DNA by incubation at 65℃ for only 1 h, and required only a simple water bath or heating block to provide a constant temperature of 65℃. LAMP amplification products had a ladder-like appearance when electrophoresed on an agarose gel. With the addition of SYBR Green Ⅰ, the presence of the target DNA could be detected by naked eyes. The detection limit of plasmid DNA (pMD19-T-VP26) by LAMP was 0.563 pg/μL which was found to be lower than that of the commonly used PCR method. The results indicate the suitability and simplicity of the test as a rapid, field diagnostic tool for WSSV.关键词
对虾白斑综合征病毒/VP26基因/环介导恒温基因扩增检测技术Key words
White spot syndrome virus(WSSV)/ VP26 gene/ Loop-mediated isothermal amplification(LAMP)引用本文复制引用
李明云,丁文超,陈炯,史雨红..环介导恒温扩增技术快速检测对虾白斑综合征病毒方法的建立[J].农业生物技术学报,2011,19(1):186-190,5.基金项目
本研究由长江学者和创新团队发展计划(No.IRT0734)、国家科技支撑计划(No.2007BAD43B0)和浙江省科技厅重点项目(No.2008C22049)共同资助 （No.IRT0734）
