Comparative analysis of PCR techniques for pathogen detection

Abstract

Polymerase chain reaction techniques have revolutionized infectious disease diagnostics. This research compared five PCR-based methods for bacterial pathogen detection: conventional PCR, real-time qPCR, multiplex PCR, digital PCR, and LAMP. Evaluation was conducted at Oceanview Institute between October 2018 and March 2020 using 450 clinical specimens containing Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii at 10 to 10⁶ CFU/mL. Digital PCR demonstrated highest sensitivity (96.3%) and specificity (99.3%) with 240-minute turnaround. Real-time qPCR achieved 92.4% sensitivity, 98.7% specificity in 90 minutes. LAMP provided fastest results (45 minutes) with 88.1% sensitivity and 97.1% specificity. These findings establish benchmarks for PCR technique selection based on clinical requirements.