Rapid and sensitive identification of pleural and peritoneal infections by droplet digital PCR
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ORIGINAL ARTICLE
Rapid and sensitive identification of pleural and peritoneal infections by droplet digital PCR Fangmei Zhou 1 & Shoudong Sun 2,3 & Xiling Sun 2 & Ye Chen 2,3 & Xuejing Yang 2 Received: 12 May 2020 / Accepted: 19 October 2020 # Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2020
Abstract 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 5fluorescent-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.
Introduction Pleural and peritoneal infections are common and important clinical problems worldwide. The incidence in both adult and pediatric populations continues to rise and causes substantial morbidity and mortality, with up to 20% of patients requiring surgery or dying (Corcoran et al. 2015). Timely and appropriate antibiotic treatment can improve the clinical outcome; however, inappropriate use and delayed initiation of antibiotics remain common. Identification of the causative organisms involved in pleural and peritoneal infections is important Fangmei Zhou and Shoudong Sun contributed equally to this work. * Xuejing Yang [email protected] 1
College of Medical Technology, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, People’s Republic of China
2
Department of Clinical Laboratory, The First Affiliated Hospital of Zhejiang Chinese Medical University, 54 Youdian Road, Hangzhou 310006, Zhejiang, People’s Republic of China
3
The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, People’s Republic of China
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