Duplex dPCR System for Rapid Identification of Gram-Negative Pathogens in the Blood of Patients with Bloodstream Infection: A Culture-Independent Approach.
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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DOI：
10.4014/jmb.2103.03044
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发表时间：
2021-11-28
影响因子：
2.8
通讯作者：
Chung YJ
中科院分区：
文献类型：
--
作者：
Shin J;Shina S;Jung SH;Park C;Cho SY;Lee DG;Chung YJ
病原体的早期和准确检测对于改善血流感染（BSI）的临床结果非常重要，特别是在耐药病原体的情况下。本研究旨在建立一种不依赖于培养的数字PCR（dPCR）系统，利用BSI患者血浆DNA进行多重检测，以检测主要败血症病原菌和耐药基因。我们的双重dPCR系统在3小时内通过5次反应成功检测到9个靶点（5个细菌特异性靶点和4个抗菌药物耐药基因）。最低检测限为50 ag细菌DNA，表明血液中1 CFU/ml的细菌可被检出。为了验证临床适用性，用我们的系统检测了发热患者的无细胞DNA样本，并证实与传统血培养的高度一致性。该系统可以支持早期鉴定一些耐药革兰阴性病原体，这有助于改善BSI的治疗结果。
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.