Development and Application of TaqMan qPCR Assays for Detecting Monkeypox Virus, Lumpy Skin Disease Virus, Goatpox Virus, Fowlpox Virus and Swinepox Virus in the Border Areas of Yunnan Province, China

The re-emergence and spread of animal poxviruses pose a significant threat to global livestock economies and public health, highlighting the need for highprecision diagnostic and surveillance tools.In this study, we developed a panel of five independent TaqMan qPCR assays utilizing adenovirus-packaged pseudovirus standards for the detection of five major animal poxviruses: monkeypox virus (MPXV), lumpy skin disease virus (LSDV), goatpox virus (GTPV), fowlpox virus (FWPV), and swinepox virus (SWPV).The constructed pseudovirus standards demonstrated good stability, with coefficients of variation (CV) below 3.46%.All five assays showed high specificity and sensitivity.The assay was further evaluated through a large-scale epidemiological survey of clinical samples collected from the transboundary high-risk border areas of Yunnan Province, China.The surveillance revealed a baseline prevalence of 13.05% for LSDV in cattle and 2.84% for SWPV in pigs.This newly established qPCR assay offers a highly sensitive, specific, and biologically safe diagnostic toolset, which may serve as a potentially useful tool for poxvirus active surveillance and transboundary animal disease control.