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KMID : 0545120210310111481
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Journal of Microbiology and Biotechnology
2021 Volume.31 No. 11 p.1481 ~ p.1489
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Duplex dPCR System for Rapid Identification of Gram-Negative Pathogens in the Blood of Patients with Bloodstream Infection: A Culture-Independent Approach
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Shin Ju-Youn
Shin Sun
Jung Seung-Hyun
Park Chul-Min
Cho Sung-Yeon
Lee Dong-Gun
Chung Yeun-Jun
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Abstract
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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 sepsiscausing 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.
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KEYWORD
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Digital PCR, bloodstream infection, Gram-negative, blood-stream infection, antimicrobial resistance
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