Primary Study on Rapid Detection of European Brown Hare Syndrome Virus by RT-LAMP

Abstract

To develop the reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) method for the detection of European brown hare syndrome virus (EBHSV), a set of 6 primers were designed according to the conserved sequence of the capsid protein(VP60)gene of EBHSV published in GenBank, and the RT-LAMP assay was described through synthesis of target DNA fragments, optimization of reaction conditions, sensitivity and specificity tests. Results showed the RT-LAMP method for the EBHSV detection had a ladder-like pattern of amplication bands from about 233 bp incubation at 64°C for 1h by using agarose gel electrophoresis, and with good specificity and sensitivity. The sensitivity of the RT-LAMP could reach about 5 copies of cloned viral genomic fragments (pGM-T-EBHSV), and no amplifications for RHDV, Pasteur Ella multocida, E.coli and Salmonella from rabbits detection by this approach.