Sensitive detection of microRNAs using polyadenine-mediated fluorescent spherical nucleic acids and a microfluidic electrokinetic signal amplification chip
- Author:
Jun XU
1
;
Qing TANG
;
Runhui ZHANG
;
Haoyi CHEN
;
Luan-Bee KHOO
;
Xinguo ZHANG
;
Yue CHEN
;
Hong YAN
;
Jincheng LI
;
Huaze SHAO
;
Lihong LIU
Author Information
1. 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
- Keywords:
MicroRNAs;
Microfluidic chip;
Electrokinetic signal amplification;
Polyadenine-DNA;
Gold nanoparticle
- From:
Journal of Pharmaceutical Analysis
2022;12(5):808-813
- CountryChina
- Language:Chinese
-
Abstract:
The identification of tumor-related microRNAs(miRNAs)exhibits excellent promise for the early diag-nosis 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 fluores-cence 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.Sub-sequently,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 re-coveries 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.
