Molecular Genetic Analysis of the Rabies Virus in Wild and Domestic Animals of Kazakhstan and the Development of a Diagnostic System

Rabies remains one of the most significant zoonotic infections, characterized by high mortality. In Kazakhstan, the virus circulates among both domestic and wild animals, necessitating ongoing molecular genetic monitoring and improved laboratory diagnostic methods. The aim of this study was to investigate the genetic characteristics of the rabies virus circulating among animals in Kazakhstan using nucleoprotein (N) gene analysis and to develop a regionally adapted real-time RT-PCR method for its detection. Kazakhstani rabies virus isolates deposited in GenBank revealed that all belong to a cosmopolitan lineage and form a single phylogenetic cluster, regardless of host species. Isolates from neighboring countries, including China and Russia, were also included in this cluster, demonstrating the regional relationship of circulating virus variants. The obtained data were used to develop a real-time PCR method based on the genetic characteristics of the rabies virus circulating in Kazakhstan. The diagnostic performance of the method was evaluated using 144 samples of various origins obtained from domestic and wild animals, including bats. The developed assay demonstrated a sensitivity of 86.4%, a specificity of 98.4%, and a diagnostic efficiency of 96.5%. The proposed method ensures reliable detection of rabies virus variants circulating in Kazakhstan and can be used for laboratory diagnostics and epidemiological monitoring.