Three-Dimensional Fluorescence Fingerprint Integrated PCR Enables Accurate Analysis of 9 Pathogens

Area of Science:

• Molecular biology
• Biotechnology
• Medical diagnostics

Background:

• Current respiratory infection diagnostics rely on culture, immunology, and PCR.
• PCR methods are sensitive but limited by signal acquisition capabilities.
• High-throughput pathogen identification is an unmet clinical need.

Purpose of the Study:

• To develop a high-throughput platform for simultaneous detection of multiple respiratory pathogens.
• To overcome signal acquisition limitations in current PCR-based diagnostics.
• To introduce 3D-FLIP9 for efficient and convenient pathogen identification.

Main Methods:

• Developed 3D-FLIP9, integrating 3D fluorescent fingerprinting with PCR.
• Enabled detection of 9 respiratory pathogens in a single tube.
• Utilized unique fluorescent fingerprints for accurate pathogen identification.

Main Results:

• Achieved high-throughput detection of 9 respiratory pathogens simultaneously.
• Demonstrated signal detectability as low as 10 copies/μL.
• Validated effectiveness in 136 clinical samples with an overall AUC of 97%.

Conclusions:

• 3D-FLIP9 offers a breakthrough in signal acquisition and processing for pathogen identification.
• The platform provides accurate identification based on unique pathogen fingerprints.
• 3D-FLIP9 has significant application value for respiratory infection prevention and control.