Application of rapid-cycle real-time polymerase chain reaction for diagnostic testing in the clinical microbiology laboratory.
BACKGROUND
Rapid-cycle real-time polymerase chain reaction (PCR) technology combines rapid thermocycling with real-time fluorescent probe detection of amplified target nucleic acids.


OBJECTIVE
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.


METHODS
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.
Cockerill
Franklin R
FR
Division of Clinical Microbiology, Mayo Medical School, Rochester, Minn 55905, USA. cockerill.franklin@mayo.edu
D001419:Bacteria / Q000235:genetics; D001424:Bacterial Infections / Q000175:diagnosis; D019411:Clinical Laboratory Techniques / Q000379:methods*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis; D014780:Viruses / Q000235:genetics