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Sensitive detection of microRNAs using polyadenine-mediated fluorescent spherical nucleic acids and a microfluidic electrokinetic signal amplification chip

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Journal of Pharmaceutical Analysis 2022年第5期12卷 808-813页

作者： Jun Xu Qing Tang Runhui Zhang Haoyi Chen Bee Luan Khoo Xinguo Zhang Yue Chen Hong Yan Jincheng Li Huaze Shao Lihong Liu NMPA Key Laboratory for Research and Evaluation of Drug Metabolism Guangdong Provincial Key Laboratory of New Drug ScreeningSchool of Pharmaceutical SciencesSouthern Medical UniversityGuangzhou510515China The Second Clinical Medical School Southern Medical UniversityGuangzhou510515China Department of Biomedical Engineering City University of Hong KongHong Kong999077China

The identification of tumor-related microRNAs(miRNAs)exhibits excellent promise for the early diagnosis of cancer and other bioanalytical applications.Therefore,we developed a sensitive and efficient biosensor using polyadenine(polyA)-mediated fluorescent spherical nucleic acid(FSNA)for miRNA analysis based on strand displacement reactions on gold nanoparticle(AuNP)surfaces and electrokinetic signal amplification(ESA)on a microfluidic chip.In this FSNA,polyA-dna biosensor was anchored on AuNP surfaces via intrinsic affinity between adenine and Au.The upright conformational polyA-dna recognition block hybridized with 6-carboxyfluorescein-labeled reporter-dna,resulting in fluorescence quenching of FSNA probes induced by AuNP-based resonance energy transfer.Reporter dna was replaced in the presence of target miRNA,leading to the recovery of reporter-dna fluorescence.Subsequently,reporter-dnas were accumulated and detected in the front of with Nafion membrane in the microchannel by ESA.Our method showed high selectivity and sensitivity with a limit of detection of 1.3 pM.This method could also be used to detect miRNA-21 in human serum and urine samples,with recoveries of 104.0%-113.3% and 104.9%-108.0%,respectively.Furthermore,we constructed a chip with three parallel channels for the simultaneous detection of multiple tumor-related miRNAs(miRNA-21,miRNA-141,and miRNA-375),which increased the detection efficiency.Our universal method can be applied to other dna/RNA analyses by altering recognition sequences.

关键词： MicroRNAs Microfluidic chip Electrokinetic signal amplification polyadenine-dna Gold nanoparticle

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