A rapid, instrument-free, sample-to-result nucleic acid amplification test

Area of Science:

• Biotechnology
• Diagnostics
• Point-of-care testing

Background:

• Current nucleic acid amplification tests (NAATs) often require specialized laboratory equipment and trained personnel.
• There is a need for portable, user-friendly diagnostic platforms for rapid pathogen detection in diverse settings.

Purpose of the Study:

• To develop and demonstrate a fully integrated, autonomous, disposable NAAT platform.
• To evaluate the performance of this platform for detecting methicillin-resistant Staphylococcus aureus (MRSA).

Main Methods:

• Development of a multiplexable autonomous disposable nucleic acid amplification test (MAD NAAT) based on two-dimensional paper networks.
• Dry reagent storage and on-device rehydration.
• Independent isothermal amplification of multiple targets with internal controls.
• Testing of human nasal swab specimens pre-screened for MRSA.

Main Results:

• The MAD NAAT platform requires no permanent instrument or manual sample processing, with a total test time under one hour.
• The prototype successfully detected MRSA in human nasal swabs.
• The lowest input copy number for consistent MRSA detection was approximately 5 × 10^3 genomic copies.

Conclusions:

• The MAD NAAT platform offers a sensitive and autonomous solution for nucleic acid testing outside traditional laboratory environments.
• This technology has the potential for widespread application in infectious disease diagnostics and surveillance.