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

Jun Xu; Qing Tang; Runhui Zhang; Haoyi Chen; Bee Luan Khoo; Xinguo Zhang; Yue Chen; Hong Yan; Jincheng Li; Huaze Shao; Lihong Liu

doi:10.1016/j.jpha.2022.05.009

Volume 12 Issue 5

Nov. 2022

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Article Navigation > Journal of Pharmaceutical Analysis > 2022 > 12(5): 808-813

Jun Xu, Qing Tang, Runhui Zhang, Haoyi Chen, Bee Luan Khoo, Xinguo Zhang, Yue Chen, Hong Yan, Jincheng Li, Huaze Shao, Lihong Liu. Sensitive detection of microRNAs using polyadenine-mediated fluorescent spherical nucleic acids and a microfluidic electrokinetic signal amplification chip[J]. Journal of Pharmaceutical Analysis, 2022, 12(5): 808-813. doi: 10.1016/j.jpha.2022.05.009

Citation:

Jun Xu, Qing Tang, Runhui Zhang, Haoyi Chen, Bee Luan Khoo, Xinguo Zhang, Yue Chen, Hong Yan, Jincheng Li, Huaze Shao, Lihong Liu. Sensitive detection of microRNAs using polyadenine-mediated fluorescent spherical nucleic acids and a microfluidic electrokinetic signal amplification chip[J]. Journal of Pharmaceutical Analysis, 2022, 12(5): 808-813. doi: 10.1016/j.jpha.2022.05.009

Citation:

Jun Xu, Qing Tang, Runhui Zhang, Haoyi Chen, Bee Luan Khoo, Xinguo Zhang, Yue Chen, Hong Yan, Jincheng Li, Huaze Shao, Lihong Liu. Sensitive detection of microRNAs using polyadenine-mediated fluorescent spherical nucleic acids and a microfluidic electrokinetic signal amplification chip[J]. Journal of Pharmaceutical Analysis, 2022, 12(5): 808-813. doi: 10.1016/j.jpha.2022.05.009

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

doi: 10.1016/j.jpha.2022.05.009

a. NMPA Key Laboratory for Research and Evaluation of Drug Metabolism, Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, China;
b. The Second Clinical Medical School, Southern Medical University, Guangzhou, 510515, China;
c. Department of Biomedical Engineering, City University of Hong Kong, Hong Kong, 999077, China

Funds:

Science and Technology Innovation Strategic Special Project of Guangdong Province ("Climbing Program" Special Project

Grant No.: pdjh2022b0106)

Guangdong College Students Innovation and Entrepreneurship Training Program (Grant No.: S202112121154).

This work was supported financially by the National Natural Science Foundation of China (Grant No.: 81973282)

National College Students Innovation and Entrepreneurship Training Program (Grant No.: 202012121024)

Guangdong Basic and Applied Basic Research Foundation (Grant Nos.: 2018A030313843 and 2021A1515011493)

Received Date: Feb. 05, 2022
Accepted Date: May 27, 2022
Rev Recd Date: May 24, 2022
Publish Date: Jun. 01, 2022

Abstract

Abstract

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.
- MicroRNAs,
- Microfluidic chip,
- Electrokinetic signal amplification,
- Polyadenine-DNA,
- Gold nanoparticle

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References(40)

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