Application of Rapid-Cycle Real-Time Polymerase Chain Reaction for Diagnostic Testing in the Clinical Microbiology Laboratory

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Abstract
Context.—Rapid-cycle real-time polymerase chain reaction (PCR) technology combines rapid thermocycling with real-time fluorescent probe detection of amplified target nucleic acids. Objectives.—To review and compare the method of rapid-cycle real-time PCR to conventional PCR methods. To describe the application of rapid-cycle real-time PCR for diagnostic testing in the microbiology laboratory. Data Selection.—Information is presented from published literature as well as from personal experience at the Mayo Clinical Microbiology Laboratory (Rochester, Minn). Conclusions.—Compared to conventional PCR methods, rapid-cycle real-time PCR diagnostics are much faster and easier to perform, and, because both PCR and probe detection occur in the same reaction vessel, the possibility of amplified product (amplicon) contamination is lessened. Furthermore, compared to conventional culture-based or direct antigen detection methods, rapid-cycle real-time PCR assays are frequently more sensitive and much more rapid techniques for detecting or quantifying microorganisms in human samples and for identifying genes or mutations in pathogens associated with antimicrobial resistance.