Ultrasensitive Competitive Lateral Flow Immunoassay with Visual Semiquantitative Inspection and Flexible Quantification Capabilities

Antibiotics abuse has caused various problems threatening human health and ecological environment. Monitoring antibiotics residual levels is of great significance, yet still challenging for quantitative point-of-need testing with high-sensitivity and visual capability. Here we developed a competitive lateral flow immunoassay (CLFIA) platform with flexible readout for enrofloxacin (ENR), a regularly added antibiotic. To overcome the limitation of low sensitivity of traditional colloidal gold-based CLFIA, the three-dimensionally assembled gold nanoparticles (AuNPs) within dendritic silica scaffold were fabricated as signal reporters. The assembly structure effectively retained the intrinsic absorption features of hydrophobic AuNPs and greatly enhanced the light extinction ability of a single label for signal amplification. The obtained CLFIA strips can not only achieve qualitative screening of ENR at a very low concentration by naked eye (cutoff value: 0.125 ng/mL), but also enable ultrasensitive quantification of ENR by an optical scanner (limit of detection: 0.00195 ng/mL) or a smartphone (limit of detection: 0.0078 ng/mL). Moreover, to elaborate the visual inspection degree of CLFIA against traditional yes/no interpretation, a novel multirange gradient CLFIA strip was prepared for visually semiquantitative identification of ENR with four concentration ranges. The novel CLFIA platform demonstrated sensitive, specific, and reliable determination of ENR with flexible signal readout and provides a potential and invigorating pathway to point-of-need immunoassay of antibiotics.