Real-time Polymerase Chain Reaction (PCR) in Microbiology

Lee, Mi Kyung

Abstract

Real-time polymerase chain reaction (PCR) is based on the detection and quantification of a fluorescent reporter. The signal increases in direct proportion to the amount of PCR product formed during each cycle in the reaction. Compared to conventional PCR methods, real-time PCR diagnostics are much faster and easier to perform, more sensitive and the possibility of amplified product contamination is lessened. Nowadays, the standard method for diagnosing the presence of pathogens in clinical samples relies on culture techniques. However, conventional method is too slow to provide clinically useful information for the physician to prescribe, only when needed, the appropriate antibiotics. More importantly, faster microbiology diagnostics appears to be an important factor influencing clinical outcome and cost reduction for healthcare. Real-time PCR assays are rapid and sensitive techniques for detecting or quantifying microorganisms and for identifying genes or mutations in pathogens associated with antimicrobial resistance. However, the accuracy of the real-time PCR data is largely depended on several factors such as sample preparation, quality of the standard and choice of housekeeping gene. Therefore, this is the major task for molecular diagnostics to guarantee reliable data in the future. New advances in real-time PCR will insure better management of patients, should reduce health costs, and could impact on the spread of antibiotic resistance.