Direct amplification of rRNA genes in diagnosis of bacterial infections.
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.
Rantakokko-Jalava
K
K
Department of Medical Microbiology, University of Turku, Turku, Finland. kaisu.rantakokko@utu.fi
Nikkari
S
S
Jalava
J
J
Eerola
E
E
Skurnik
M
M
Meurman
O
O
Ruuskanen
O
O
Alanen
A
A
Kotilainen
E
E
Toivanen
P
P
Kotilainen
P
P
D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D001419:Bacteria / Q000302:isolation & purification; D001424:Bacterial Infections / Q000175:diagnosis* / Q000188:drug therapy; D001431:Bacteriological Techniques; D016208:Databases, Factual; D005387:Finland; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification*; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000302:isolation & purification; D012338:RNA, Ribosomal, 23S / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA
D000900:Anti-Bacterial Agents; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S