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Rapid and sensitive identification of pleural and peritoneal infections by droplet digital PCR
Fangmei Zhou1, Shoudong Sun2,3, Xiling Sun2
1College of Medical Technology, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, People's Republic of China.
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 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.
Insights
A novel droplet digital PCR (ddPCR) assay offers rapid and sensitive detection of pathogens in pleural and peritoneal infections. This culture-independent method significantly reduces diagnostic time compared to traditional methods.
Area of Science:
- Infectious Diseases
- Molecular Diagnostics
- Microbiology
Background:
- Pleural and peritoneal infections are serious conditions associated with high morbidity and mortality.
- Current diagnostic methods, primarily based on microbial culture, are time-consuming and lack sensitivity.
- Rapid and accurate pathogen identification is crucial for effective treatment and patient outcomes.
Purpose of the Study:
- To evaluate a 5-fluorescent-channel droplet digital PCR (ddPCR) system for the rapid, culture-independent detection of pathogens in pleural and peritoneal fluid.
- To compare the performance of ddPCR with traditional culture-based methods for diagnosing these infections.
- To assess the sensitivity, specificity, and turnaround time of the ddPCR assay.
Main Methods:
- A 5-fluorescent-channel droplet digital PCR (ddPCR) system with 5 assay panels was utilized.
- Pleural fluid (n=40) and peritoneal fluid (n=19) samples were analyzed using ddPCR.
- Results were compared against traditional culture methods performed on the same samples.
Main Results:
- The ddPCR assay demonstrated high sensitivity: 96% for pleural fluid and 92.86% for peritoneal fluid.
- ddPCR detected more polymicrobial infections (11 in pleural, 3 in peritoneal) than culture (4 in pleural, 2 in peritoneal).
- The turnaround time for ddPCR was approximately 3 hours, significantly faster than the 38.30 ± 22.44 hours for culture.
Conclusions:
- The ddPCR assay is a rapid, sensitive, and promising method for identifying pathogens in pleural and peritoneal infections.
- This approach facilitates earlier diagnosis and has the potential to optimize patient treatment strategies.
- ddPCR offers a significant advancement over traditional culture-based diagnostics for these critical infections.
