Nucleic acid amplification tests for Yersinia pestis detection: A comprehensive technical and field-oriented review

Plague, caused by Yersinia pestis, remains a highly fatal zoonosis with endemic foci on three continents. Rapid and accurate diagnosis is essential for patient survival and outbreak containment. Nucleic acid amplification tests (NAATs) have evolved from conventional PCR to real-time PCR, isothermal methods (LAMP, RPA), droplet digital PCR, and CRISPR-based platforms. This 5000-word review provides an in-depth technical comparison of these technologies, their analytical performance (limit of detection [LoD], sensitivity, specificity), clinical validation in human specimens, environmental and vector applications, and point-of-care (POC) readiness. We critically analyze sample preparation challenges, genetic target selection, and multiplexing strategies. Four comprehensive tables summarize target genes, real-time PCR assays, isothermal methods, and point-of-care platforms. Future directions include AI-integrated devices, wearable sensors, and lyophilized CRISPR tests. This review serves as a definitive guide for clinical microbiologists, field epidemiologists, and diagnostic developers.