Multiplex PCR in diagnosis and characterization of bovine viral diarrhoea virus isolates from India.

摘要

Though prevalence of bovine viral diarrhoea virus (BVDV) antibodies in Indian cattle has been established by serology, information on the use of multiplex PCR for diagnosis and characterization of BVDV isolates at the genomic level is lacking. This study developed a multiplex polymerase chain reaction (PCR) assay using primers of 5' untranslated region (UTR) and structural glycoprotein (E1-E2) region, which generated two different amplicons (288 bp and 784 bp) in a single tube when 13 Indian BVDV isolates were tested. Both the amplicons were found specific when restriction enzyme analysis and subsequent nucleotide sequencing of three selected isolates representing different geographic areas of India were performed. The sequence analysis of both the regions grouped them into BVDV 1b genotype. The results demonstrated that multiplex PCR can be used for identification and subsequent genotyping of BVDV isolates.