Surface Decorated Magnetic Particles in Combination with Isothermal Nucleic Acid Amplification for Rapid Pathogen Detection

Abstract

Nucleic acid extraction from clinical samples followed by nucleic acid amplification tests newline(NAATs) is the key step of conventional molecular diagnostics. Recently, isothermal nucleic newlineacid amplification tests (iNAATs) draw the attention of scientists. These techniques have a newlinesignificant advantage over conventional NAATs as the detection of the amplicons by iNAATs newlinecan be pushed forward using a thermal cycler-free manner, therefore enabling field detection. newlineIt helps obtain rapid, quantitative results either in the laboratory or, more importantly, in a newlineresource-limited area. Despite these advantages, prior nucleic acid extraction from the clinical newlinesample is necessary for both NAATs and iNAATs, which need high-cost instruments, a newlinecentralized lab, and highly trained personnel. Microfluidic devices can overcome the issue of newlineprior nucleic acid extraction but the complex fabrication procedure of the analytical device newlinemakes it non-compatible in the resource-constrained settings by the end-users. Immunoassays newlinecould also be applied to detect the viable pathogen, but the high cost and uncertainty of newlineantibodies are still limitations for those methods. Keeping in mind these research gaps, our newlineprimary research is mainly focused on surface-modified magnetic particle-mediated nucleic newlineacid extraction from infectious pathogen followed by downstream isothermal amplification newlinefor near-point-of-care diagnostics, especially for the resource-constrained area. Our second newlinearea of research explores the alternatives to costly antigen-based detection methods by newlineemploying aptamers to detect viable pathogen. Our first objective explores two chitosancoated newlinemagnetic particle preparation methods that can be executed within 6 hours from newlinecommonly available chemicals with nothing but a magnetic stirrer and water bath. It is also newlinedoable by a minimally trained person followed by downstream loop-mediated isothermal newlineamplification (LAMP).