Simultaneous assay for ten bacteria and toxins in spiked clinical samples using a microflow cytometer

Area of Science:

• Biomedical Engineering
• Clinical Diagnostics
• Microfluidics

Background:

• Bacterial infections and intoxications share common symptoms, necessitating rapid diagnostic tools.
• Accurate identification of pathogens and toxins is vital for timely and effective treatment.
• Previous microflow cytometry methods could analyze six targets in buffer.

Purpose of the Study:

• To expand microflow cytometry multiplexing capabilities to ten bacteria and toxins.
• To validate multiplexed immunoassays in complex clinical matrices like serum and nasal wash.
• To assess the performance of microflow cytometry against established antibody-based detection methods.

Main Methods:

• Multiplexed immunoassays were developed and performed using a microflow cytometer.
• Assays were tested with ten different bacteria and toxins.
• Performance was evaluated in buffer, spiked clinical samples (serum, nasal wash), and blind clinical trials, incorporating positive and negative controls.

Main Results:

• Microflow cytometry achieved detection limits comparable to or superior to ELISA, lateral flow, and array biosensors.
• Detection in complex matrices (serum, nasal wash) yielded limits of detection similar to those in buffer samples.
• Successful analysis of spiked clinical samples in blind trials confirmed assay robustness.

Conclusions:

• Microflow cytometry is a powerful platform for multiplexed detection of bacteria and toxins.
• The enhanced method demonstrates high sensitivity and accuracy in complex clinical samples.
• This technology offers a promising avenue for rapid and comprehensive pathogen diagnostics.