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Establishment of a RT-lamp method for the rabbit hemorrhagic disease virus detection

Yang Zexiao, Zeng Hui, Wang Yin, Yao Xueping, Wang Kaiyu

To develop a new molecular biological method for the quick detection of rabbit hemorrhagic disease virus (RHDV), a set of 7 primerswere designed according to the conserved sequence of the capsid proteinVP60gene of RHDV published inGenBank, and the reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was established through preparation of target gene fragments, optimization of reaction conditions, sensitivity and specificity tests. Results showed the RT-LAMPmethod for the RHDV detection had a ladder-like pattern of amplication bands from about 213 bp incubation at 64ºC for 45min by using agarose gel electrophoresis,and with good sensitivity and specificity,the detection limit could reach about 5 copies of cloned viral genomic fragments, which was more sensitve than that of traditional RT-PCR, and no amplifications for gene fragment of European brown hare syndrome virus, Pasteurella multocida, E.coli and Salmonella fromrabbits detection by this RT-LAMP approach.