A paper and plastic device for performing recombinase polymerase amplification of HIV DNA

Area of Science:

• Biotechnology
• Medical Diagnostics
• Public Health

Background:

• Early diagnosis of HIV in infants is crucial but challenging in low- and middle-income countries.
• Current gold standard DNA PCR testing is resource-intensive and lacks point-of-care accessibility.
• A significant gap exists in accessible, rapid HIV DNA testing for early infant diagnosis.

Purpose of the Study:

• To develop an inexpensive, portable device for isothermal amplification of HIV DNA.
• To enable point-of-care HIV DNA testing in resource-limited settings.
• To facilitate early infant diagnosis of HIV through accessible technology.

Main Methods:

• Development of a multi-layered paper, glass fiber, and plastic device for isothermal enzymatic amplification.
• Incorporation of lyophilized enzymes and facilitated reaction component mixing within the device.
• Utilized recombinase polymerase amplification (RPA) in a five-step operational process.
• Detection of amplified HIV DNA using commercially available lateral flow strips.

Main Results:

• The device successfully amplified 10 copies of HIV DNA to detectable levels within 15 minutes.
• Demonstrated the feasibility of using the device with dried-blood spots after DNA extraction.
• The device is inexpensive, lightweight, and easy to assemble, suitable for low-resource environments.

Conclusions:

• The developed paper-based device shows promise for point-of-care HIV DNA testing.
• This technology could significantly improve early infant diagnosis of HIV in resource-limited settings.
• Integration with lateral flow strips offers a viable solution for accessible HIV diagnostics.