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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, Jari Jalava,^1, 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.

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^* 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.

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

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

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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.

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