Advances in rapid microbiological testing for animal diseases: A review

Animal pathogenic microorganisms destabilize livestock economies and jeopardize human health through zoonotic transmission and foodborne illness. Traditional culture-based detection methods, while standardized, are time-consuming and labor-intensive, often failing to meet the urgent need for rapid on-site or point-of-care (POC) monitoring required to prevent disease outbreaks and manage animal health effectively. By integrating latest research advances, this study reviews advances in rapid detection technologies for animal pathogens, including the evolution of nucleic acid amplification strategies, with a focused comparison of the analytical sensitivity and field deployability of quantitative polymerase chain reaction (qPCR) and mainstream isothermal amplification techniques (loop-mediated isothermal amplification (LAMP); recombinase polymerase amplification (RPA)). Furthermore, this study reports on the emergence of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated protein (Cas) systems as next-generation diagnostic tools, highlighting their integration with microfluidic Lab-on-a-Chip (LOC) platforms to achieve attomolar sensitivity. We also consider the application of portable nanopore sequencing for real-time pathogen identification from clinical livestock samples and the growing role of Artificial Intelligence (AI) in analyzing complex diagnostic datasets. Advanced molecular methods have achieved significant reductions in time consumption from days to less than one hour while challenges regarding sample preparation from complex clinical matrices such as whole blood, serum, tissue homogenates, and fecal samples remain. The future of animal health surveillance lies in integrated, automated systems that combine the specificity of CRISPR-Cas diagnostics with the connectivity of IoT-enabled biosensors for farm-level early warning. Comparative analysis indicates that isothermal amplification methods (LAMP, RPA) coupled with CRISPR-Cas systems offer the optimal balance of sensitivity, speed, and field deployability for POC veterinary diagnostics, while qPCR/dPCR (dPCR)remain indispensable for quantitative regulatory applications such as disease certification and vaccine efficacy monitoring. We propose a structured technology selection framework to guide researchers and veterinary practitioners in choosing appropriate detection modalities based on specific sensitivity, cost, throughput, and deployment requirements for different livestock species and production systems.