Lab-on-a-Film disposable for genotyping multidrug-resistant Mycobacterium tuberculosis from sputum extracts

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Microfluidics

Background:

• Multidrug-resistant tuberculosis (MDR-TB) poses a significant global health challenge, necessitating rapid and accessible diagnostic tools.
• Current diagnostic methods can be costly and time-consuming, limiting their widespread use in resource-limited settings.
• Development of innovative, low-cost disposable diagnostic platforms is crucial for effective MDR-TB management.

Purpose of the Study:

• To introduce and characterize a novel Lab-on-a-Film disposable device for the detection of multidrug-resistant tuberculosis (MDR-TB).
• To evaluate the analytical sensitivity and specificity of the Lab-on-a-Film system for identifying key MDR-TB mutations.
• To demonstrate the utility of the Lab-on-a-Film device in detecting MDR-TB from clinical sputum specimens.

Main Methods:

• Development of a Lab-on-a-Film disposable featuring 203 gel elements with DNA probes for 37 MDR-TB mutations across 5 genes.
• Integration of multiplex amplification and hybridization within a single microfluidic flow cell on a flexible, low-cost film.
• Utilization of a closed-amplicon system with integrated post-hybridization wash steps to prevent cross-contamination.
• Automated sample extraction using a TruTip workstation followed by amplification and detection on the Lab-on-a-Film disposable.

Main Results:

• The Lab-on-a-Film disposable demonstrated an analytical sensitivity of 32 colony-forming units per milliliter (cfu mL-1) for all MDR-TB markers.
• The system successfully detected MDR-TB in positive clinical sputum specimens.
• The cost of the gel elements on the flexible film is approximately 500 times less than traditional microarray glass.

Conclusions:

• The Lab-on-a-Film disposable represents a cost-effective and sensitive platform for MDR-TB detection.
• The integrated microfluidic design simplifies the diagnostic workflow, minimizing contamination risks.
• This technology has the potential to enhance MDR-TB diagnostics, particularly in settings with limited resources.