Optimisation of PCR conditions for the detection of infectious bovine rhinotracheitis

The relevance of this study is determined by the need to improve the efficiency of laboratory diagnostics of persistent viral infections in cattle, which are characterised by low pathogen concentrations and difficulty in detection using traditional methods. The aim of the study was to develop and optimise a polymerase chain reaction (PCR) for accurate identification of bovine infectious rhinotracheitis virus based on analysis of the gB gene. Modern methods of molecular biology, genetic engineering, and bioinformatics analysis were used, including the collection, processing, and alignment of nucleotide sequences. During the study, nucleotide sequences of the gB gene of bovine infectious rhinotracheitis virus were collected and analysed, which allowed the identification of its conserved, variable, and polymorphic regions. Based on the results of the analysis, a highly specific pair of primers was designed and synthesised to ensure selective amplification of the target viral genome fragment. DNA of the bovine infectious rhinotracheitis virus was isolated to optimise amplification conditions. Optimal temperature parameters of the polymerase chain reaction and the quantitative composition of reagents were also determined, ensuring high sensitivity and specificity of the method. The results confirmed that the efficiency of diagnostics can be improved through rational selection of primers and reaction parameters. The practical value of this work is that the obtained data and developed methodological approaches can be used by veterinary diagnostic laboratories and research institutions to improve PCR diagnostics of bovine infectious rhinotracheitis virus