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  • Title: Rapid and sensitive detection of Singapore grouper iridovirus by loop-mediated isothermal amplification.
    Author: Mao XL, Zhou S, Xu D, Gong J, Cui HC, Qin QW.
    Journal: J Appl Microbiol; 2008 Aug; 105(2):389-97. PubMed ID: 18312563.
    Abstract:
    AIMS: 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. 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 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. CONCLUSIONS: 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. 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.
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