One-Step Surface Modification to Graft DNA Codes on Paper: The Method, Mechanism, and Its Application

Glass slides have been widely used for DNA immobilization in DNA microarray and numerous bioassays for decades, whereas they are faced with limitations of low probe density, time-consuming modification steps, and expensive instruments. In this work, a simple one-step surface modification method using 3-aminopropyl trimethoxysilane (APTMS) has been developed and applied to graft DNA codes on paper. Higher DNA immobilization efficiency was obtained in comparison with that in a conventional method using glass slides. Fluorescence detection, X-ray photo-electron spectroscopy (XPS), infrared spectra (FT-IR), and pH influence studies were employed to characterize the surface modification and subsequent DNA immobilization, which further reveals a mechanism in which this method lies in ionic interactions between the positively charged APTMS-modified paper surface and negatively charged DNA probes. Furthermore, an APTMS-modified paper-based device has been developed to demonstrate application in low-cost detection of a foodborne pathogen, Giardia lamblia, with high sensitivity (the detection limit of 22 nM) and high specificity. Compared with conventional methods using redundant cross-linking reactions, our method is simpler, faster, versatile, and lower-cost, enabling broad applications of paper-based bioassays especially for point-of-care detection in resource-poor settings.