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Rapid and sensitive identification of pleural and peritoneal infections by droplet digital PCR
Folia Microbiologica ( IF 2.6 ) Pub Date : 2020-11-07 , DOI: 10.1007/s12223-020-00834-0
Fangmei Zhou 1 , Shoudong Sun 2, 3 , Xiling Sun 2 , Ye Chen 2, 3 , Xuejing Yang 2
Affiliation  

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.



中文翻译:

通过液滴数字 PCR 快速、灵敏地识别胸膜和腹膜感染

胸膜和腹膜感染导致大量的发病率和死亡率。传统的诊断方法依赖于临床样本的培养,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,我们评估了一个 5 荧光通道液滴数字 PCR (ddPCR) 系统和 5 个测定板,用于直接从胸膜和腹膜液样本中进行独立于培养的快速病原体检测。同一样品的传统培养物被用作参考。本研究共检测了 40 个胸水样品和 19 个腹腔液体样品。胸水标本检测阳性25例,培养4例多菌感染,ddPCR阳性26例,多菌感染11例;肺炎克雷伯菌是胸膜和腹膜液样本中最常见的细菌。胸水和腹膜液样本的 ddPCR 检测灵敏度分别为 96%(95% 置信区间 (CI) = 79.65 至 99.90%)和 92.86%(95% CI = 66.13 至 99.82%)。ddPCR 检测的周转时间约为 3 小时,而基于培养的鉴定为 38.30 ± 22.44 小时。我们的结果表明,ddPCR 检测是一种快速、灵敏的方法,可用于识别导致胸膜和腹膜感染的病原体,并且将成为早期诊断和优化感染治疗的有前途的方法。

更新日期:2020-11-09
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