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1. Rapid and Sensitive Detection of Singapore grouper iridovirus by Loop-Mediated Isothermal Amplification | |||
Mao Xinliang,Zhou Sheng,Xu Dan,Gong Jie,Cui Huachun,Qin Qiwei ,Qin Qiwei | |||
Biology 24 December 2007 | |||
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Abstract:Aims: The aim of this paper is to develop a loop-mediated isothermal amplification (LAMP) method for rapid, sensitive and inexpensive detection of SGIV in grouper, Epinephelus sp. Methods and Results: 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°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 7 other aquatic animal viruses indicating high specificity of the LAMP. The LAMP method was applied to detect SGIV in virus infected GP cells and grouper tissues effectively. Conclusions: The LAMP described in this study is a cheap, sensitive, specific and rapid protocol for detection of SGIV in cells and in grouper tissues. Significance and Impact of the Study: The developed LAMP method can be simply applied both in field condition and in laboratory operation for specific detection of SGIV infection. | |||
TO cite this article:Mao Xinliang,Zhou Sheng,Xu Dan, et al. Rapid and Sensitive Detection of Singapore grouper iridovirus by Loop-Mediated Isothermal Amplification[OL].[24 December 2007] http://en.paper.edu.cn/en_releasepaper/content/17288 |
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