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Journal of Clinical Microbiology, October 2001, p. 3446-3451, Vol. 39, No. 10
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.10.3446-3451.2001

Sensitive and Specific Method for Rapid Identification of Streptococcus pneumoniae Using Real-Time Fluorescence PCR

James C. McAvin,1 Patricia A. Reilly,2 Robert M. Roudabush,1 William J. Barnes,1 Ann Salmen,3 Glen W. Jackson,3 Kathleen K. Beninga,3 Alicia Astorga,1 Ferne K. McCleskey,3 William B. Huff,4 Debra Niemeyer,5 and Kenton L. Lohman1,*

Molecular Epidemiology Branch,1 Clinical Microbiology Branch,3 and Epidemiology Surveillance Division,4 Air Force Institute for Environment and Occupational Health Risk Analysis/Epidemiology Surveillance Division, Brooks Air Force Base, San Antonio, and Clinical Microbiology, Wilford Hall Medical Center, Lackland Air Force Base, Lackland,2 Texas, and Medical NBC Sciences and Technology Directorate, Office of the Air Force Surgeon General, Bolling Air Force Base, Washington, D.C.5

Received 22 March 2001/Returned for modification 16 May 2001/Accepted 18 July 2001

Molecular surveillance of pathogens has shown the need for rapid and dependable methods for the identification of organisms of clinical and epidemiological importance. As the leading cause of community-acquired pneumonia, Streptococcus pneumoniae was used as a model organism to develop and refine a real-time fluorescence PCR assay and enhanced DNA purification method. Seventy clinical isolates of S. pneumoniae, verified by latex agglutination, were screened against 26 negative control clinical isolates employing a TaqMan assay on a thermocycler (LightCycler). The probe, constructed from the lytA gene, correctly detected all S. pneumoniae genomes without cross-reaction to negative controls. The speed and ease of this approach will make it adaptable to identification of many bacterial pathogens and provide potential for adaptation to direct detection from patient specimens.

* Corresponding author. Mailing address: Molecular Epidemiology, AFIERA, 2601 Westgate Rd., Building 930, Brooks AFB, San Antonio, TX 78235. Phone: (210) 536-2639. Fax: (210) 536-2638. E-mail: kenton.lohman{at}brooks.af.mil.

Journal of Clinical Microbiology, October 2001, p. 3446-3451, Vol. 39, No. 10
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.10.3446-3451.2001


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