Rapid DNA visual detection of polymicrobial bloodstream infection using filter paper

Area of Science:

• Biomedical Diagnostics
• Materials Science
• Molecular Biology

Background:

• Bloodstream infections (BSI) are a significant complication in cancer patients, increasing mortality and healthcare costs.
• Current blood culture methods for BSI diagnosis are slow and often miss polymicrobial infections, which are common and severe in neutropenic patients.
• Effective antimicrobial management requires rapid and accurate pathogen identification.

Purpose of the Study:

• To develop a rapid diagnostic tool for simultaneous detection of multiple pathogens causing BSI in neutropenic cancer patients.
• To address the limitations of conventional blood cultures in identifying polymicrobial BSI.
• To enable timely and targeted antimicrobial therapy.

Main Methods:

• Fabrication of a functionalized cellulose filter paper surface using glutaric anhydride, N-hydroxysuccinimide, and N,N'-dicyclohexylcarbodiimide.
• Development of a triplex DNA detection assay targeting common BSI pathogens: Escherichia coli, Saccharomyces cerevisiae, and human cytomegalovirus.
• Evaluation of detection time and sensitivity of the functionalized paper assay.

Main Results:

• The functionalized paper enabled rapid detection of all three target pathogen DNAs within 1-5 minutes.
• The assay demonstrated a low detection limit of 0.1-0.5 ng/µL for the targeted pathogens.
• Successful identification of triplex pathogen DNA was achieved.

Conclusions:

• The developed functionalized paper assay offers a potential solution for rapid polymicrobial BSI diagnosis.
• This tool can support timely and accurate antimicrobial treatment decisions for high-risk patients.
• Integration into portable, automated equipment could further enhance clinical utility.