Duplex dPCR system for rapid identification of gram-negative pathogens in the blood of patients with bloodstream infection: A c

Shin, J., Shina, S., Jung, S. H., Park, C., Cho, S. Y., Lee, D. G., & Chung, Y. J. (2021). Duplex dPCR system for rapid identification of gram-negative pathogens in the blood of patients with bloodstream infection: A culture-independent approach. J Microbiol Biotechnol, 31(1).

DOI: 10.4014/jmb.2103.03044

Abstract

Early and accurate detection of pathogens is important to improve clinical outcomes of bloodstream infections (BSI), especially in the case of drug-resistant pathogens. In this study, we aimed to develop a culture-independent digital PCR (dPCR) system for multiplex detection of major sepsis-causing gram-negative pathogens and antimicrobial resistance genes using plasma DNA from BSI patients. Our duplex dPCR system successfully detected nine targets (five bacteria-specific targets and four antimicrobial resistance genes) through five reactions within 3 hours. The minimum detection limit was 50 ag of bacterial DNA, suggesting that 1 CFU/ml of bacteria in the blood can be detected. To validate the clinical applicability, cell-free DNA samples from febrile patients were tested with our system and confirmed high consistency with conventional blood culture. This system can support early identification of some drug-resistant gram-negative pathogens, which can help improving treatment outcomes of BSI.