Sensitive glucometer-based microfluidic immune-sensing platform via DNA signal amplification coupled with enzymatic reaction

•A novel glucometers-based microfluidic immune-biosensor for protein biomarker detection was developed for the first time.•A 43 × 43 mm microfluidic chip with small reagent volumes which can be directly mass-manufactured were prepared.•This biosensor didn’t require professional equipment except a glucose meter and can provide a higher detection efficiency.•This strategy has universal significance for applications in sensing other biomarkers by changing antibody or aptamers. Here we present a novel glucometer-based microfluidic platform using miniaturized personal glucose meter (PGM) with portability and simple operation to sensitively detect cancer biomarker via amplification of rolling cycle amplification (RCA) associated with DNA-invertase catalysis. This design utilizes gold nanoparticles (Au NPs) as a supporting matrix to anchor Human Epidermal Growth Factor Receptor 2 antibody (anti-HER2) and DNA sequence that is able to induce RCA. After adding HER2, a sandwiches immunocomplex forms between a peptide and the asymmetric DNA-anti-HER2-modified Au NPs in the detection area of a glass slide, followed by RCA process and hybridization of the RCA products with DNA-invertase for enzymatic generation of glucose. In such a way, RCA-amplified DNA converted glucose can be detected by PGM for sensitively diagnosis of HER2 biomarker. This assay is performed on a 43 × 43 mm microfluidic chip with small reagent volume, large dynamic range and high sensitivity, holding great promise in the clinical diagnosis at home or in urban area.