A LAMP-based microfluidic module for rapid detection of pathogen in cryptococcal meningitis.

Area of Science:

• Medical Diagnostics
• Biotechnology
• Infectious Diseases

Background:

• Cryptococcal meningitis (CM) poses a significant global health burden, characterized by high morbidity and mortality.
• Current diagnostic methods for CM are limited, particularly concerning rapid and accessible detection technologies.
• Microfluidic approaches for CM diagnosis are still in early stages of development and require further investigation.

Purpose of the Study:

• To develop a multifunctional microfluidic module for integrated pathogen enrichment and on-chip nucleic acid extraction for CM diagnosis.
• To create a simplified, rapid, and efficient diagnostic approach for Cryptococcus in cerebrospinal fluid (CSF) samples.
• To assess the potential of the microfluidic module as a replacement for or adjunct to existing diagnostic methods like LFA.

Main Methods:

• Development of a microfluidic module incorporating a filtration membrane for Cryptococcus cell capture and on-chip nucleic acid extraction.
• Optimization of nucleic acid extraction using lyticase digestion, alkaline lysis, and heating within the microfluidic module.
• Integration of a portable, one-pot, lyophilized LAMP reagent bead for temperature-independent operation.

Main Results:

• The microfluidic module successfully integrated pathogen enrichment and nucleic acid extraction, reducing sample processing risks.
• A portable, instrument-free system was developed, enabling a 'sample in and answer out' detection strategy for real CSF samples.
• The developed module shows promise for replacing LFA in later stages of CM treatment and as an auxiliary diagnostic tool in early stages.

Conclusions:

• The developed multifunctional microfluidic module offers a simple, rapid, and efficient approach for diagnosing cryptococcal meningitis.
• This technology has practical prospects for improving CM diagnosis and management, particularly in resource-limited settings.
• The microfluidic module is a promising tool for both early diagnosis confirmation and monitoring treatment efficacy in CM.