Electrochemical detection of miRNA-222 by use of a magnetic bead-based bioassay

被引:112
|
作者
Bettazzi, Francesca [1 ]
Hamid-Asl, Ezat [1 ,4 ]
Esposito, Carla Lucia [2 ]
Quintavalle, Cristina [3 ]
Formisano, Nello [3 ]
Laschi, Serena [1 ]
Catuogno, Silvia [2 ,3 ]
Iaboni, Margherita [3 ]
Marrazza, Giovanna [1 ]
Mascini, Marco [1 ]
Cerchia, Laura [2 ]
De Franciscis, Vittorio [2 ]
Condorelli, Gerolama [2 ,3 ]
Palchetti, Ilaria [1 ]
机构
[1] Univ Florence, Dipartimento Chim Ugo Schiff, I-50019 Sesto Fiorentino, Fi, Italy
[2] CNR, Ist Endocrinol & Oncol Sperimentale, I-80138 Naples, Italy
[3] Univ Naples Federico II, Dipartimento Biol & Patol Cellulare & Mol, I-80138 Naples, Italy
[4] Univ Mazandran, Fac Chem, Dept Analyt Chem, Babol Sar, Iran
关键词
MicroRNA; Magnetic beads; Electrochemical assay; miRNA-222; MICRORNA SIGNATURES; TRAIL RESISTANCE; DNA; BIOSENSOR; CANCER; ACID; LNA;
D O I
10.1007/s00216-012-6476-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
MicroRNAs (miRNAs, miRs) are naturally occurring small RNAs (approximately 22 nucleotides in length) that have critical functions in a variety of biological processes, including tumorigenesis. They are an important target for detection technology for future medical diagnostics. In this paper we report an electrochemical method for miRNA detection based on paramagnetic beads and enzyme amplification. In particular, miR 222 was chosen as model sequence, because of its involvement in brain, lung, and liver cancers. The proposed bioassay is based on biotinylated DNA capture probes immobilized on streptavidin-coated paramagnetic beads. Total RNA was extracted from the cell sample, enriched for small RNA, biotinylated, and then hybridized with the capture probe on the beads. The beads were then incubated with streptavidin-alkaline phosphatase and exposed to the appropriate enzymatic substrate. The product of the enzymatic reaction was electrochemically monitored. The assay was finally tested with a compact microfluidic device which enables multiplexed analysis of eight different samples with a detection limit of 7 pmol L-1 and RSD = 15 %. RNA samples from non-small-cell lung cancer and glioblastoma cell lines were also analyzed.
引用
收藏
页码:1025 / 1034
页数:10
相关论文
共 50 条
  • [1] Electrochemical detection of miRNA-222 by use of a magnetic bead-based bioassay
    Francesca Bettazzi
    Ezat Hamid-Asl
    Carla Lucia Esposito
    Cristina Quintavalle
    Nello Formisano
    Serena Laschi
    Silvia Catuogno
    Margherita Iaboni
    Giovanna Marrazza
    Marco Mascini
    Laura Cerchia
    Vittorio De Franciscis
    Gerolama Condorelli
    Ilaria Palchetti
    Analytical and Bioanalytical Chemistry, 2013, 405 : 1025 - 1034
  • [2] Magnetic bead-based hybridization assay for electrochemical detection of microRNA
    Bartosik, Martin
    Hrstka, Roman
    Palecek, Emil
    Vojtesek, Borivoj
    ANALYTICA CHIMICA ACTA, 2014, 813 : 35 - 40
  • [3] Magnetic bead-based hybridization assay for electrochemical detection of microRNA
    Bartosik, Martin (martin.bartosik@mou.cz), 1600, Elsevier B.V., Netherlands (813):
  • [4] Magnetic bead-based hybridization assay for electrochemical detection of microRNA
    Bartosik, M. (martin.bartosik@mou.cz), 1600, Elsevier B.V., Netherlands (813):
  • [5] Magnetic bead-based label-free electrochemical detection of DNA hybridization
    Wang, J
    Kawde, AN
    Erdem, A
    Salazar, M
    ANALYST, 2001, 126 (11) : 2020 - 2024
  • [6] Fluorescent magnetic bead-based mast cell biosensor for electrochemical detection of allergens in foodstuffs
    Jiang, Donglei
    Zhu, Pei
    Jiang, Hui
    Ji, Jian
    Sun, Xiulan
    Gu, Wenshu
    Zhang, Genyi
    BIOSENSORS & BIOELECTRONICS, 2015, 70 : 482 - 490
  • [7] Multifunctional magnetic bead-based electrochemical immunoassay for the detection of aflatoxin B1 in food
    Tang, Dianping
    Zhong, Zhaoyang
    Niessner, Reinhard
    Knopp, Dietmar
    ANALYST, 2009, 134 (08) : 1554 - 1560
  • [8] Single bead-based electrochemical biosensor
    Liu, Changchun
    Schrlau, Michael G.
    Bau, Haim H.
    BIOSENSORS & BIOELECTRONICS, 2009, 25 (04): : 809 - 814
  • [9] Magnetic bead-based electrochemical assay for determination of DNA methyltransferase activity
    Dudova, Zdenka
    Bartosik, Martin
    Fojta, Miroslav
    ELECTROCHIMICA ACTA, 2017, 231 : 575 - 581
  • [10] Bead-based immunoassays: Fluorescence vs. electrochemical detection.
    Farrell, S
    Ronkainen-Matsuno, NJ
    Halsall, HB
    Heineman, WR
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2002, 224 : U126 - U127