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Journal of Clinical Microbiology, January 2000, p. 32-39, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Direct Amplification of rRNA Genes in Diagnosis of Bacterial Infections

Kaisu Rantakokko-Jalava,1,* Simo Nikkari,1,2,dagger Jari Jalava,1,Dagger Erkki Eerola,1 Mikael Skurnik,3 Olli Meurman,2 Olli Ruuskanen,4 Anna Alanen,5 Esa Kotilainen,6 Paavo Toivanen,1 and Pirkko Kotilainen7

Departments of Medical Microbiology,1 Clinical Microbiology,2 Medical Biochemistry,3 Pediatrics,4 Obstetrics and Gynecology,5 Surgery,6 and Medicine,7 University of Turku, Turku, Finland

Received 6 July 1999/Returned for modification 28 August 1999/Accepted 22 September 1999

A broad-range bacterial PCR targeting rRNA genes (rDNAs) was used to directly analyze 536 clinical samples obtained from 459 hospitalized patients during a 4-year study period. The molecular diagnosis based on DNA sequencing of the PCR product was compared to that obtained by bacterial culture. The bacteriological diagnosis was concordant for 447 (83%) specimens. Broad-range rDNA PCR was the only method that yielded an etiologic diagnosis for 11 (2.4%) of 459 patients. Compared to culture and clinical assessment, the sensitivity of the PCR method combined with sequencing was 74.2%, and the specificity was between 98.7 and 99.6%. At present, the described molecular approach proved superior to bacterial culture in two clinical situations: infections caused by bacteria with unusual growth requirements and specimens taken during antimicrobial treatment of the patient.

* Corresponding author. Mailing address: Department of Medical Microbiology, University of Turku, Kiinamyllynkatu 13, 20520 Turku, Finland. Phone: 358-2-3337423. Fax: 358-2-2330008. E-mail: kaisu.rantakokko{at}utu.fi.

dagger Present address: Department of Microbiology and Immunology, Stanford University School of Medicine, VA Palo Alto Health Care System 154T, Palo Alto, CA 94304.

Dagger Present address: National Public Health Institute, 20520 Turku, Finland.

Journal of Clinical Microbiology, January 2000, p. 32-39, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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