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Clinical Microbiology Reviews, January 2006, p. 165-256, Vol. 19, No. 1
0893-8512/06/$08.00+0     doi:10.1128/CMR.19.1.165-256.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

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 III, and T. F. Smith

Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota

Real-time PCR has revolutionized the way clinical microbiology laboratories diagnose many human microbial infections. This testing method combines PCR chemistry with fluorescent probe detection of amplified product in the same reaction vessel. In general, both PCR and amplified product detection are completed in an hour or less, which is considerably faster than conventional PCR detection methods. Real-time PCR assays provide sensitivity and specificity equivalent to that of conventional PCR combined with Southern blot analysis, and since amplification and detection steps are performed in the same closed vessel, the risk of releasing amplified nucleic acids into the environment is negligible. The combination of excellent sensitivity and specificity, low contamination risk, and speed has made real-time PCR technology an appealing alternative to culture- or immunoassay-based testing methods for diagnosing many infectious diseases. This review focuses on the application of real-time PCR in the clinical microbiology laboratory.

* Corresponding author. Mailing address: Mayo Clinic, 200 First St. SW, Hilton 470, Rochester, MN 55905. Phone: (507) 284-4682. Fax: (507) 284-4272. E-mail: espy.mark{at}mayo.edu.

Clinical Microbiology Reviews, January 2006, p. 165-256, Vol. 19, No. 1
0893-8512/06/$08.00+0     doi:10.1128/CMR.19.1.165-256.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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