Evaluation of 16S rRNA gene PCR and culture of buffy coat, 16srRNA gene PCR of whole blood in comparison with whole blood culture for detection of bacteremia

• Main Authors: Valle, Demetrio L., Jr., Andrade, Jeannie I., Cabrera, Esperanza C., Rivera, Windell L.
• Format: text
• Published: Animo Repository 2022
• Subjects: Bacteremia; Blood—Buffy coat; Leucocytes; Polymerase chain reaction; Pathogenic bacteria; Biology; Pathological Conditions, Signs and Symptoms
• Online Access: https://animorepository.dlsu.edu.ph/faculty_research/5088
• Institution: De La Salle University

The use of Gram-type specific PCR on buffy coat from clinical specimens for the detection of bacteremia 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 ten 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 comparable to blood culture. The Kappa coefficients of agreement were 0.9819 (buffy coat PCR), 0.9458 (whole blood PCR), and 1.0 (buffy coat culture), establishing their validity as alternative methods for the routine blood culture in detecting bacteremia. In addition, results showed that there was a direct correlation of WBC counts greater than 12,000 cells per mm3 to the occurrence of bacteremia as detected by the four methods (P <0.05)