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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 GreenⅠ). 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.
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Received: 20 March 2010
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