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Rapid detection of Macrobrachium rosenbergii nodavirus ( Mr NV) and extra small virus (XSV), the pathogenic agents of white tail disease of Macrobrachium rosenbergii (De Man), by loop‐mediated isothermal amplification
Author(s) -
Pillai D,
Bonami JR,
Sri Widada J.
Publication year - 2006
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/j.1365-2761.2006.00718.x
Subject(s) - macrobrachium rosenbergii , loop mediated isothermal amplification , biology , agarose gel electrophoresis , virology , prawn , virus , white spot syndrome , shrimp , agarose , microbiology and biotechnology , fishery , dna , genetics
A loop‐mediated isothermal amplification (LAMP) procedure is described for rapid diagnosis of white tail disease, a viral disease caused by Macrobrachium rosenbergii nodavirus ( Mr NV) and extra small virus (XSV), in the giant freshwater prawn, Macrobrachium rosenbergii . This method was more sensitive than conventional RT‐PCR for detecting the two viruses. A set of four primers, two outer and two inner, were designed for Mr NV detection. An additional pair of loop primers was also used in an accelerated LAMP reaction for detection of XSV. Time and temperature conditions were optimized for detection of the two viruses. The LAMP reaction is highly suited for disease diagnosis in developing countries as amplification of DNA can be detected without the use of agarose gel electrophoresis, by the production of whitish precipitate of magnesium pyrophosphate as a by‐product.