Express indication of pathogens of viral diarrhea, infectious rhinotracheitis, parainfluenza-3 in cattle

Introduction. In recent years, pathologies of the digestive and respiratory systems have often been recorded among cattle. The leading etiological factors of respiratory diseases are respiratory syncytial virus, viruses of parainfluenza and infectious rhinotracheitis. Respiratory viral infections cause great economic damage to dairy cattle breeding Considering the mixed course of viral respiratory infections, the development of simultaneous amplification of pathogen genomes specific fragments is of particular interest. Purpose. Simultaneous detection of genomes of pathogens of viral diarrhea, infectious rhinotracheitis and parainfluenza-3 by PCR in real time. Materials and methods. In this study, cultures of bovine embryonic kidney cells (BEK), bovine kidney cell line (NDBK) artificially infected with viruses, as well as biomaterial were used. For the detection of the genomes of the VD, IRT, and PI viruses, the most conservative sections of the viral genomes were selected: the genome fragment with the length of 4716−4846 bp in the HV genome for IRT agent, genome fragment with the length of 84−204 bp for VD agent, and genome fragment with the length of 2820−2910 bp for PI agent. To control amplification in Real-time mode (positive control), the design and synthesis of terminal plasmid DNA molecules containing amplifiable DNA sequences was performed. Research results, discussion. Samples from sick animals and pathological material were tested using the multiplex PCR in Real-time mode. The results obtained showed the high specificity and efficiency of the multiplex polymerase chain reaction for the indication and identification of pathogens. Conclusion. The developed set and mode of Real-time polymerase chain reaction allowed us to determine simultaneously 3 different genomes of causative agents of respiratory infections in cattle.