Limited-resource preparable chitosan magnetic particles for extracting amplification-ready nucleic acid from complex...

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Materials Science

Background:

• Nucleic acid extraction is crucial for pathogen detection and genetic disease diagnosis but conventional methods are resource-intensive.
• Existing lab-on-a-chip and magnetic bead methods have limitations including cost, complexity, and reliance on hazardous chemicals.
• There is a need for simple, instrument-free nucleic acid extraction methods suitable for limited-resource settings.

Purpose of the Study:

• To investigate chitosan-coated magnetic particles for instrument-free nucleic acid extraction.
• To assess the efficiency and suitability of these particles for downstream nucleic acid amplification tests (NAATs).
• To develop a method for decentralized nucleic acid-based diagnostics in resource-limited environments.

Main Methods:

• Developed easily preparable chitosan-coated magnetic particles using basic lab equipment (water bath, magnetic stirrer).
• Quantitatively assessed DNA magnetocapture efficiency using UV260.
• Extracted DNA from various samples (aqueous solution, cell lysate, serum) and detected it using quantitative real-time loop-mediated isothermal amplification (LAMP) and real-time polymerase chain reaction (PCR).

Main Results:

• Chitosan magnetic particles efficiently captured and released nucleic acids.
• Extracted DNA from 100-1000 copies of *Escherichia coli* or human genomic DNA was successfully detected by LAMP and PCR.
• The entire process, from magnetocapture to NAAT, had a turnaround time of 1.5-2 hours.

Conclusions:

• Chitosan-coated magnetic particles offer a simple, instrument-free method for nucleic acid extraction.
• The method is compatible with sensitive downstream NAATs and suitable for limited-resource settings.
• This technology can facilitate the expansion and decentralization of nucleic acid-based diagnostics globally.