用于检测大肠杆菌血流感染患者病原体的数字液滴PCR的临床诊断性能：一项前瞻性观察研究
Clinical diagnostic performance of droplet digital PCR for pathogen detection in patients with Escherichia coli bloodstream infection: a prospective observational study.
作者信息
Kitagawa Hiroki, Kojima Masato, Tadera Kayoko, Kogasaki Shuta, Omori Keitaro, Nomura Toshihito, Shigemoto Norifumi, Hiyama Eiso, Ohge Hiroki
机构信息
Department of Infectious Diseases, Hiroshima University Hospital, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan.
Department of Surgery, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan.
出版信息
BMC Infect Dis. 2025 Jan 6;25(1):22. doi: 10.1186/s12879-024-10396-y.
BACKGROUND
Droplet digital PCR (ddPCR) is a highly sensitive tool for detecting bacterial DNA in bacterial bloodstream infections (BSI). This study aimed to examine the sensitivity and specificity of ddPCR and the association between bacterial DNA load in whole blood and the time-to-positivity (TTP) of blood culture (BC) in patients with Escherichia coli BSI.
METHODS
This prospective study enrolled patients with E. coli BSI confirmed via BC at the Hiroshima University Hospital from June 2023 to August 2024. The E. coli DNA load in whole blood, which was simultaneously obtained from two BC sets, was measured using ddPCR with E. coli specific primer and probe. Whole blood samples from 50 patients with BC positive for pathogens other than E. coli (n = 25) and BC negative (n = 25) were also evaluated using ddPCR.
RESULTS
A total of 131 patient samples were analyzed in this study. Of the 81 patients with E. coli BSI, ddPCR detected E. coli DNA in 67 (82.7%). The results of ddPCR for E. coli had a sensitivity of 82.7% (95% CI: 73.1-89.4%), specificity 100% (95% CI: 93.0-100%). Patients with positive ddPCR results had significantly shorter TTP than those with negative results (median, 8.8 h vs. 10.7 h, p < 0.001). The positivity rate for both BC sets was significantly higher in patients with positive ddPCR results than in those with negative results (89.6% vs. 35.1%, p < 0.001). Among ddPCR-positive patients, septic shock was significantly associated with intestinal perforation, higher E. coli DNA load, higher 28-d mortality, shorter TTP, and higher positivity rate for four bottles of BC than those without septic shock. The E. coli DNA load in whole blood negatively correlated with TTP (p < 0.001, R = 0.38).
CONCLUSION
The E. coli DNA load in whole blood is inversely correlated with TTP. Notably, a higher E. coli DNA load is associated with septic shock.
背景
液滴数字PCR（ddPCR）是检测细菌血流感染（BSI）中细菌DNA的高灵敏度工具。本研究旨在探讨ddPCR的敏感性和特异性，以及大肠杆菌BSI患者全血中细菌DNA载量与血培养（BC）阳性时间（TTP）之间的关联。
方法
这项前瞻性研究纳入了2023年6月至2024年8月在广岛大学医院通过血培养确诊为大肠杆菌BSI的患者。使用针对大肠杆菌的特异性引物和探针，通过ddPCR检测同时从两套血培养瓶中获取的全血中的大肠杆菌DNA载量。还使用ddPCR评估了50例血培养结果为除大肠杆菌外其他病原体阳性（n = 25）和血培养阴性（n = 25）患者的全血样本。
结果
本研究共分析了131份患者样本。在81例大肠杆菌BSI患者中，ddPCR检测到67例（82.7%）存在大肠杆菌DNA。ddPCR检测大肠杆菌的结果敏感性为82.7%（95%CI：73.1 - 89.4%），特异性为100%（95%CI：93.0 - 100%）。ddPCR结果阳性的患者TTP显著短于结果阴性的患者（中位数，8.8小时对10.7小时，p < 0.001）。ddPCR结果阳性的患者两套血培养瓶的阳性率显著高于结果阴性的患者（89.6%对35.1%，p < 0.001）。在ddPCR阳性患者中，感染性休克与肠穿孔、较高的大肠杆菌DNA载量、较高的28天死亡率、较短的TTP以及四瓶血培养较高的阳性率显著相关，高于无感染性休克的患者。全血中大肠杆菌DNA载量与TTP呈负相关（p < 0.001，R = 0.38）。
结论
全血中大肠杆菌DNA载量与TTP呈负相关。值得注意的是，较高的大肠杆菌DNA载量与感染性休克相关。
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