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

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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 * #

College of Life Science, Sun Yat-sen University

*Correspondence author

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Funding: “973”计划课题,“863”计划,教育部博士点基金,教育部博士点基金,广东省科技计划项目（No.2006CB101802,2006AA100306,20040558011,20050558043,2006B50104003）

Opened online:24 December 2007

Accepted by: none

Citation: Mao Xinliang,Zhou Sheng,Xu Dan.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

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.

Keywords:SGIV; LAMP; Grouper; Nested PCR

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