Franklin R Cockerill1. 1. Division of Clinical Microbiology, Mayo Medical School, Rochester, Minn 55905, USA. cockerill.franklin@mayo.edu
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.
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.
Authors: M J Espy; J R Uhl; L M Sloan; S P Buckwalter; M F Jones; E A Vetter; J D C Yao; N L Wengenack; J E Rosenblatt; F R Cockerill; T F Smith Journal: Clin Microbiol Rev Date: 2006-01 Impact factor: 26.132
Authors: Mark A Poritz; Anne J Blaschke; Carrie L Byington; Lindsay Meyers; Kody Nilsson; David E Jones; Stephanie A Thatcher; Thomas Robbins; Beth Lingenfelter; Elizabeth Amiott; Amy Herbener; Judy Daly; Steven F Dobrowolski; David H-F Teng; Kirk M Ririe Journal: PLoS One Date: 2011-10-19 Impact factor: 3.240
Authors: Marcela Agne Alves Valones; Rafael Lima Guimarães; Lucas André Cavalcanti Brandão; Paulo Roberto Eleutério de Souza; Alessandra de Albuquerque Tavares Carvalho; Sergio Crovela Journal: Braz J Microbiol Date: 2009-03-01 Impact factor: 2.476