Analytica chimica acta · 2026
Design and fabrication of a dual-mode colorimetric nanobiosensor for detection of breast cancer-related microRNA biomarkers upon the simultaneous application of gold nanoparticles and DNAzyme.
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Keramati Maryam, Rahaie Mahdi
Аннотация
BACKGROUND: Breast cancer is among the most prevalent malignancies worldwide, and early diagnosis is essential to improve treatment efficacy. In this study, we developed a dual signal colorimetric nanobiosensor for the simultaneous detection of two breast cancer-related microRNAs, miR-145 and miR-21.
METHODS: The nanobiosensor utilizes target-triggered hybridization chain reaction (HCR) to form G-quadruplex DNAzymes for miR-145 detection via OPD oxidation (at 425 nm). For miR-21, target-induced hybridization leads to aggregation of gold nanoparticles (AuNPs) and the absorbance peak at 535 nm. In multiplexed analysis, miR-145 contributes to both signals, while miR-21 specifically affects the 535 nm absorbance, enabling simultaneous quantification.
RESULTS: The nanobiosensor demonstrated excellent sensitivity and specificity for detecting target microRNAs, achieving limits of detection (LOD) as low 0.15 nM for miR-145 and 0.12 nM for miR-21. Strong linear correlations were observed, with R2 values of 0.9887 for miR-145 and 0.9711 for miR-21. The nanobiosensor effectively distinguished target microRNAs both individually and simultaneously, while miR-195 and miR-155 were used as non-targets to validate its high specificity.
SIGNIFICANCE: In experiments involving the co-detection of both targets, the nanobiosensor achieved LODs in the range of 0.01-0.4 nM, wich further confirmed the nanobiosensor's stability and efficient multiplex detection capability. In conclusion, the developed dual-miRNA detection platform offers a powerful assay for the early screening of breast cancer and demonstrates strong potential for application in clinical multiplex diagnostic assays.
METHODS: The nanobiosensor utilizes target-triggered hybridization chain reaction (HCR) to form G-quadruplex DNAzymes for miR-145 detection via OPD oxidation (at 425 nm). For miR-21, target-induced hybridization leads to aggregation of gold nanoparticles (AuNPs) and the absorbance peak at 535 nm. In multiplexed analysis, miR-145 contributes to both signals, while miR-21 specifically affects the 535 nm absorbance, enabling simultaneous quantification.
RESULTS: The nanobiosensor demonstrated excellent sensitivity and specificity for detecting target microRNAs, achieving limits of detection (LOD) as low 0.15 nM for miR-145 and 0.12 nM for miR-21. Strong linear correlations were observed, with R2 values of 0.9887 for miR-145 and 0.9711 for miR-21. The nanobiosensor effectively distinguished target microRNAs both individually and simultaneously, while miR-195 and miR-155 were used as non-targets to validate its high specificity.
SIGNIFICANCE: In experiments involving the co-detection of both targets, the nanobiosensor achieved LODs in the range of 0.01-0.4 nM, wich further confirmed the nanobiosensor's stability and efficient multiplex detection capability. In conclusion, the developed dual-miRNA detection platform offers a powerful assay for the early screening of breast cancer and demonstrates strong potential for application in clinical multiplex diagnostic assays.