Towards Comprehensive Molecular Investigation of Rotavirus A Prevalence in Neonatal Calves and Pregnant Cows

Searching for the presence of species Rotavirus A, or Group A rotaviruses (RVA), in clinical samples could be essential for confirming diagnosis on the etiologic agent, and Reverse Transcription PCR (RT-PCR) amplification of the virus’ capsid VP6 gene in animal feces is the first choice as the protein is considered as the determinant of species. For that purpose, we have applied several adopted primer pairs with fecal samples from local calves and cows and obtained mostly negative outcomes. The formation non-specific “trash” bands of PCR products has prompted us to check capacity of the pairs in amplifying the species-specific gene. By screening the sameness/conservativeness of VP6 gene sequences of RVA bovine/cow strains available in GenBank Database we have chosen several fragments that can serve as consensus primers for detection of almost all of the already sequenced strains of the species. The theoretical outcomes were much increased, meanwhile the RT-PCR products of fecal samples proved that the pair of primers worked well in amplification of the target fragment of the gene. Thus, the capacity of VP6 gene-amplifying RT-PCR could be broadened for more comprehensive epidemiological investigation of the viral infections.