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Published on: March 26, 2013
Intraoperative pathogen rapid detection on chip
Quanying Fu1, Zhikai Wu2, Shaoqiong Feng1
1Guangdong Provincial Key Laboratory of Sensing Technology and Biomedical Instrument, School of Biomedical Engineering, Shenzhen Campus of Sun Yat-sen University, Shenzhen, 518107, China.
Abstract:
During surgery, purulent fluid containing pathogens may be found at the surgical site. Pathogens in purulent fluid can cause sepsis if disseminated into the bloodstream. Therefore, intraoperative detection of pathogens in purulent fluid is critical for guiding surgeons in selecting surgical management and antibiotic therapy. However, current detection methods are either time-consuming or unable to identify live pathogens, restricting their clinical application in intraoperative settings. To overcome these limitations, we proposed an intraoperative pathogen rapid detection (IPRD) method. This method can simultaneously detect the presence, viable fraction, species, and concentrations of multiple pathogens in 15 min. This method is performed on a chip with two parts: a single-channel part combines live/dead staining with AI-assisted microscopy to quantify live pathogens, and a parallel-channel part uses electroporation-based lysis followed by LAMP assay for nucleic acid detection. This method demonstrated an accuracy of 99.01 % in clinical validation, reliably detecting Candida albicans, Escherichia coli, and Enterococcus faecalis at 103 CFU/mL. The IPRD method has potential applications in various infection-prone surgeries, offering timely pathogenic information to assist surgeons in making surgical decisions.
Insights
A new intraoperative pathogen rapid detection (IPRD) method identifies live pathogens in surgical fluid within 15 minutes. This rapid diagnostic tool aids surgeons in timely treatment decisions for surgical site infections.
Area of Science:
- Medical Diagnostics
- Microbiology
- Surgical Innovation
Background:
- Surgical site infections are a significant risk, potentially leading to sepsis.
- Accurate and rapid pathogen identification intraoperatively is crucial for effective treatment.
- Current methods for pathogen detection are often too slow or cannot distinguish live from dead organisms.
Purpose of the Study:
- To develop a rapid, chip-based method for intraoperative pathogen detection.
- To simultaneously identify pathogen presence, viability, species, and concentration.
- To provide timely information for surgical and antibiotic therapy decisions.
Main Methods:
- A novel intraoperative pathogen rapid detection (IPRD) method was developed.
- The method utilizes a chip with two parts: live/dead staining with AI microscopy and electroporation-based lysis followed by LAMP assay.
- Simultaneous detection of multiple pathogens, viability, and concentration in 15 minutes.
Main Results:
- The IPRD method achieved 99.01% accuracy in clinical validation.
- It reliably detected Candida albicans, Escherichia coli, and Enterococcus faecalis at 10^3 CFU/mL.
- The assay provides comprehensive pathogenic information within a clinically relevant timeframe.
Conclusions:
- The IPRD method offers a significant advancement in intraoperative diagnostics.
- It enables rapid and accurate identification of live pathogens in surgical fluid.
- This technology has potential applications in infection-prone surgeries, improving patient outcomes.
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