College of Medical Technology, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, People's Republic of China.
Department of Clinical Laboratory, The First Affiliated Hospital of Zhejiang Chinese Medical University, 54 Youdian Road, Hangzhou, 310006, Zhejiang, People's Republic of China.
Folia Microbiol (Praha). 2021 Apr;66(2):213-219. doi: 10.1007/s12223-020-00834-0. Epub 2020 Nov 7.
Pleural and peritoneal infections cause substantial morbidity and mortality. Traditional diagnostic methods rely on the cultivation of clinical samples, which usually takes days to obtain report and holds a low detection sensitivity. In this study, we evaluated a 5-fluorescent-channel droplet digital PCR (ddPCR) system and 5 assay panels for culture-independent rapid pathogen detections directly from pleural and peritoneal fluid samples. Traditional culture of the same sample was used as reference. A total of 40 pleural fluid samples and 19 peritoneal fluid samples were tested in this study. Twenty-five positives including 4 polymicrobial infections by culture and 26 positives including 11 polymicrobial infections by ddPCR were detected for pleural fluid samples; 14 positives including 2 polymicrobial infections by culture and 15 positives including 3 polymicrobial infections by ddPCR were detected for peritoneal fluid samples. Klebsiella pneumoniae was the most common bacterium detected both in pleural and in peritoneal fluid samples. The sensitivity of the ddPCR assay for pleural and peritoneal fluid samples was 96% (95% confidence interval (CI) = 79.65 to 99.90%) and 92.86% (95% CI = 66.13 to 99.82%), respectively. The turnaround time of the ddPCR assay was approximately 3 h comparing with 38.30 ± 22.44 h for culture-based identifications. Our results demonstrated that the ddPCR assay is a rapid and sensitive method for identifying pathogens responsible for pleural and peritoneal infections and would be a promising approach for early diagnosis and optimizing treatment of infections.
胸膜和腹膜感染会导致严重的发病率和死亡率。传统的诊断方法依赖于临床样本的培养，这通常需要数天才能获得报告，并且检测灵敏度较低。在这项研究中，我们评估了一种 5 荧光通道液滴数字 PCR（ddPCR）系统和 5 个用于直接从胸膜和腹膜液样本中进行非培养快速病原体检测的检测面板。传统的相同样本培养作为参考。本研究共检测了 40 份胸膜液样本和 19 份腹膜液样本。对于胸膜液样本，检测到 25 个阳性结果，包括 4 个培养的混合感染和 26 个 ddPCR 的阳性结果，包括 11 个混合感染；对于腹膜液样本，检测到 14 个阳性结果，包括 2 个培养的混合感染和 15 个 ddPCR 的阳性结果，包括 3 个混合感染。肺炎克雷伯菌是在胸膜和腹膜液样本中最常见的细菌。ddPCR 检测方法对胸膜和腹膜液样本的敏感性分别为 96%（95%置信区间（CI）=79.65 至 99.90%）和 92.86%（95% CI=66.13 至 99.82%）。ddPCR 检测方法的周转时间约为 3 小时，而基于培养的鉴定需要 38.30±22.44 小时。我们的结果表明，ddPCR 检测方法是一种快速、敏感的方法，可用于鉴定导致胸膜和腹膜感染的病原体，有望为感染的早期诊断和优化治疗提供一种有前途的方法。
