Detection of porcine parvovirus by loop-mediated isothermal amplification.

Loop-mediated isothermal amplification is a novel method for rapid amplification of DNA. It has been adopted widely for the detection of virus because of its simplicity, rapidity, and specificity. A loop-mediated isothermal amplification assay was developed for the detection of porcine parvovirus. Four primers specific for six regions of PPV non-structural protein 1 gene were designed with an online software. After amplifying at a constant temperature of 59-65 degrees C by Bst enzyme, a clear result was visible after 2.5% agarose gel electrophoresis. The sensitivity and specificity of this assay were evaluated by comparison with the polymerase chain reaction. The detection limit of the assay was shown to be equivalent to 5 PPV copies/reaction. Due to its specificity and simplicity, the assay should be a useful diagnostic tool for epidemiologic studies of PPV.