An integrated dual functional recognition/amplification bio-label for the one-step impedimetric detection of Micro-RNA-21

被引:51
作者
Azzouzi, Sawsen [1 ,2 ]
Mak, Wing Cheung [1 ]
Kor, Kamalodin [1 ,3 ]
Turner, Anthony P. F. [1 ]
Ben Ali, Mounir
Beni, Valerio [1 ,4 ]
机构
[1] Linkoping Univ, Biosensors & Bioelect Ctr, Dept Phys Chem & Biol IFM, S-58183 Linkoping, Sweden
[2] Univ Sousse, Higher Inst Appl Sci & Technol Sousse, GREENS ISSAT, Cite Ettafala, Ibn Khaldoun Sousse 4003, Tunisia
[3] Iranian Natl Inst Oceanog & Atmospher Sci, Tehran 141554781, Iran
[4] ACREO SWEDISH ICE AB, S-60117 Norrkoping, Sweden
关键词
MicroRNA-21; Gold nanoparticles; Molecular beacon; Electrochemical impedance spectroscopy; GOLD NANOPARTICLES; SENSITIVE DETECTION; MOLECULAR BEACONS; GUIDED FORMATION; MICRORNAS; BIOSENSOR; SERUM; ASSAY; EXPRESSION; CANCER;
D O I
10.1016/j.bios.2017.02.014
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Alteration in expression of miRNAs has been correlated with different cancer types, tumour stage and response to treatments. In this context, a structurally responsive oligonucleotide-based electrochemical impedimetric biosensor has been developed for the simple and sensitive detection of miRNA-21. A highly specific biotinylated DNA/LNA molecular beacon (MB) probe was conjugated with gold nanoparticles (AuNPs) to create an integrated, dual function bio-label (biotin-MB-AuNPs) for both biorecognition and signal generation. In the presence of target miRNA-21, hybridisation takes place resulting in the "activation" of the biotin-MB; this event makes the biotin group, which was previously "protected" by the steric hindrance of the MB stem-loop structure, accessible. The activated biotin-MB-AuNPs/miRNA complexes become available for capture, via supramolecular interaction, onto a nentravidin-modified electrode for electrochemical transduction. The binding event results in a decrease of the charge transfer resistance at the working electrode/electrolyte interface. The biosensor responded linearly in the range 1-1000 pM of miRNA-21, with a limit of detection of 0.3 pM, good reproducibility (Relative Standard deviation (RSD) =3.3%) and high selectivity over other miRNAs (i.e. miRNA221 and miRNA-205) sequences. Detection of miRNA-21 in spiked serum samples at clinically relevant levels (low pM range) was also demonstrated, thus illustrating the potential of the biosensor for point-of-care clinical applications. The proposed biosensor design, based on the combination of a neutravidin transducing surface and the dual-function biotin-MB-AuNPs bio-label, provides a simple and robust approach for detection of short-length nucleic acid targets, such as miRNAs.
引用
收藏
页码:154 / 161
页数:8
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