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

Area of Science:

• Medical Microbiology
• Molecular Diagnostics
• Zoonotic Diseases

Background:

• Plague, caused by Yersinia pestis, is a severe zoonotic disease with global foci.
• Accurate and rapid diagnostics are crucial for patient outcomes and controlling outbreaks.
• Nucleic acid amplification tests (NAATs) are key diagnostic tools for plague.

Purpose of the Study:

• To provide a comprehensive technical comparison of various NAATs for Yersinia pestis detection.
• To evaluate the analytical performance, clinical validation, and point-of-care readiness of different NAAT platforms.
• To identify challenges and future directions in plague diagnostics.

Main Methods:

• Review of conventional PCR, real-time PCR, isothermal methods (LAMP, RPA), droplet digital PCR, and CRISPR-based platforms.
• Analysis of analytical performance metrics including limit of detection, sensitivity, and specificity.
• Evaluation of clinical validation in human specimens and applications in environmental and vector surveillance.

Main Results:

• Detailed comparison of NAAT technologies, highlighting their strengths and limitations.
• Summary of performance data and clinical utility across different diagnostic platforms.
• Identification of key challenges in sample preparation, target selection, and multiplexing.

Conclusions:

• NAATs have significantly advanced plague diagnostics, offering diverse options for various settings.
• Point-of-care readiness varies among platforms, with ongoing development towards more accessible solutions.
• Future advancements may involve AI integration and novel formats like lyophilized CRISPR tests for improved plague detection and management.