Development of a RT-PCR for the Detection of European Brown Hare Syndrome Virus
To develop the reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of European brown hare syndrome virus (EBHSV), 3 specific primers and 9 overlapping oligo primers were designed to amplify a fragment of 335 nucleotides from the genome of EBHSV according to the genome sequences published in GenBank, and a 359 bp DNA fragment of the capsid protein(VP60)gene of EBHSV was synthesized in vitro by using overlap extension PCR to construct the recombinant plasmids pGM-T-EBHSV, then the RT-PCR method was described through the optimization of reaction conditions, sensitivity and specificity tests. Results showed the RT-PCR method was supplied for the detection of EBHSV with good specificity and sensitivity. The sensitivity of the RT-PCR could reach about 50 copies of cloned viral genomic fragments (pGM-T-EBHSV), and no amplifications for RHDV, Pasteurella multocida, E.coli and Salmonella from rabbits detection by this approach.
Y. Z. Xiao et al., "Development of a RT-PCR for the Detection of European Brown Hare Syndrome Virus", Advanced Materials Research, Vols. 271-273, pp. 410-416, 2011