Exploring a phage-based real-time PCR assay for diagnosing Acinetobacter baumannii bloodstream infections with high sensitivity

摘要

In the present study, we developed a phage-based real-time quantitative PCR (qPCR) methodology for sensitive diagnosis of bloodstream infection (BSI) caused by Acinetobacter baumannii (A. baumannii). An isolated A. baumannii phage p53 was used for Taqman qPCR through detecting phage replication in live A. baumannii cells in serum samples. At the phage concentration of 10(3) PFU/mL, the sensitive detection of A. baumannii (down to 10 CFU in 100 mu L serum) has been obtained within 4 h in spiked serum samples without bacteria isolation and DNA extraction. Subsequent testing of 22 simulated serum samples spiked by different strains has shown that the results from the phage-based Taqman qPCR method have 100% agreement with the spiked concentrations of the bacteria. The assay built in this study, gathering all the advantages for detections of high rapidity, high sensitivity, good specificity, being able to detect only live bacteria not dead bacteria and no DNA extraction or purifications, can be developed to detecting other bacterial pathogens in serum or other complicated samples through switching to other types of phages and realize the rapid and sensitive detection of bacteria in BSI, which would potentially be applied for fast diagnosis in sepsis clinically. (C) 2018 Elsevier B.V. All rights reserved.