title: Evaluation of buffy coat 16S rRNA PCR, buffy coat culture and whole blood PCR for detection of bacteraemia.
abstract: The use of Gram type-specific PCR on buffy coat from clinical specimens for the detection of bacteraemia was evaluated for the first time using whole blood culture as the gold standard. In addition, the established buffy coat culture and whole blood PCR were also compared. Gram-positive bacteria belonging to six species and Gram-negative bacteria from 10 species were isolated and identified by culture and detected using broad-range 16S rDNA primers and Gram-specific primers. Data from the three methods all conferred very high sensitivity, specificity, positive and negative predictive values when compared to whole blood culture. The Kappa coefficients of agreement were 0.9819 (buffy coat PCR), 0.9458 (whole blood PCR) and 1.0 (buffy coat culture), which establishes their validity as alternative methods to routine blood culture in detecting bacteraemia. In addition, results showed that there was a direct correlation of WBC counts greater than 12,000 cells per mm(3) to the occurrence of bacteraemia as detected by the four methods (p < 0.05).
authors.affiliation: Department of Laboratories, Ospital ng Makati, Makati City, Philippines.
authors.forename: Demetrio L
authors.initials: DL
authors.lastname: Valle
authors.forename: Jeannie I
authors.initials: JI
authors.lastname: Andrade
authors.forename: Esperanza C
authors.initials: EC
authors.lastname: Cabrera
authors.forename: Windell L
authors.initials: WL
authors.lastname: Rivera
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D016470:Bacteremia / Q000175:diagnosis* / Q000382:microbiology; D046508:Culture Techniques / Q000379:methods; D005260:Female; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000302:isolation & purification*; D006094:Gram-Positive Bacteria / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007962:Leukocytes / Q000382:microbiology*; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D012336:RNA, Ribosomal, 16S