Paper-Origami-Based Multiplexed Malaria Diagnostics from Whole Blood.

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Point-of-Care Testing

Background:

• Accurate and rapid microbial species identification is crucial for effective disease management.
• Current diagnostic methods can be time-consuming, expensive, and require specialized laboratory equipment.
• There is a need for accessible, field-deployable diagnostic tools for infectious diseases.

Purpose of the Study:

• To develop and validate a novel, integrated paper-based origami device for the multiplexed determination of microbial species.
• To enable rapid, sample-to-answer nucleic acid-based diagnostics from a small volume of whole blood.
• To assess the diagnostic accuracy of the developed method for Plasmodium species detection compared to PCR.

Main Methods:

• Utilized a paper-folding origami technique for sequential DNA extraction, loop-mediated isothermal amplification (LAMP), and array-based fluorescence detection.
• Employed a low-cost handheld flashlight for naked-eye detection of DNA amplicons.
• Analyzed 80 patient blood samples, comparing results against the gold-standard polymerase chain reaction (PCR) assay in an operator-blinded study.

Main Results:

• Successfully demonstrated multiplexed microbial species determination from whole blood using the integrated origami device.
• Achieved 'sample-to-answer' diagnosis within 45 minutes from a finger-prick blood volume with minimal user intervention.
• Identified three Plasmodium species with high diagnostic accuracy, comparable to PCR, even with stored or fixed samples.

Conclusions:

• The paper-folding origami device offers a rapid, low-cost, and user-friendly platform for point-of-care microbial diagnostics.
• This technology has significant potential for the early and accurate diagnosis of infectious diseases, such as malaria, in resource-limited settings.
• The method's robustness with various sample types supports its applicability in diverse clinical scenarios.