Mao X L, Zhou S, Xu D, Gong J, Cui H C, Qin Q W
State Key Laboratory of Biocontrol, College of Life Science, Sun Yat-sen University, Guangzhou, PR China.
J Appl Microbiol. 2008 Aug;105(2):389-97. doi: 10.1111/j.1365-2672.2008.03761.x. Epub 2008 Feb 29.
The aim of this paper was to develop a loop-mediated isothermal amplification (LAMP) method for rapid, sensitive and inexpensive detection of Singapore grouper iridovirus (SGIV) in grouper (GP), Epinephelus sp.
A set of six specific primers was designed by targeting the SGIV ORF-014L. With Bst DNA polymerase large fragment, the target DNA can be amplified as early as 20 min at 65 degrees C in a simple water bath. The detection limit is about 0.02 fg (equivalent to 6.3 copies) of plasmid ORF-014L. LAMP products could be judged with three different methods. There were no cross-reactions with seven other aquatic animal viruses indicating high specificity of the LAMP. The LAMP method was applied to detect SGIV in virus-infected GP cells and GP tissues effectively.
The LAMP described in this study is a cheap, sensitive, specific and rapid protocol for the detection of SGIV in cells and in GP tissues.
The developed LAMP method can be simply applied both in field condition and in laboratory operation for specific detection of SGIV infection.
本文旨在开发一种环介导等温扩增（LAMP）方法，用于快速、灵敏且廉价地检测石斑鱼（Epinephelus sp.）中的新加坡石斑鱼虹彩病毒（SGIV）。
通过靶向SGIV的ORF-014L设计了一组六条特异性引物。使用Bst DNA聚合酶大片段，在简单的水浴中于65℃下最早20分钟即可扩增目标DNA。检测限约为0.02 fg（相当于6.3个拷贝）的质粒ORF-014L。LAMP产物可用三种不同方法判断。与其他七种水生动物病毒无交叉反应，表明LAMP具有高度特异性。LAMP方法有效地应用于检测病毒感染的石斑鱼细胞和石斑鱼组织中的SGIV。
本研究中描述的LAMP是一种用于检测细胞和石斑鱼组织中SGIV的廉价、灵敏、特异且快速的方法。
所开发的LAMP方法可简单地应用于现场条件和实验室操作中，用于SGIV感染的特异性检测。
