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ADAM, V. HÚSKA, D. HUBÁLEK, J. KIZEK, R.
Original Title
Easy to use and rapid isolation and detection of a viral nucleic acid by using paramagnetic microparticles and carbon nanotubes-based screen-printed electrodes
Type
journal article - other
Language
English
Original Abstract
A method that is easy to use, rapid, with a low cost of detecting viral nucleic acid in a biological sample represents the essential tool in targeted therapy. In this study, we report the use of paramagnetic microparticles covered by streptavidin and modified by an oligonucleotide probe with a specific viral sequence labeled by biotin to detect human immunodeficiency virus (HIV) and influenza virus subtype H5N1. The viral nucleic acids were primarily detected by adsorptive transfer stripping technique coupled with square wave voltammetry using carbon paste, hanging mercury drop or carbon nanotubes-based screen-printed working electrodes. Detection limits were estimated for both sequences down to picograms per 3 mu l. To isolate the viral sequences, paramagnetic microparticles covered with biotin-labeled oligonucleotides were used. We calculated the yield of isolation for H5N1 and/or HIV sequences, which was defined as "isolated concentration of viral nucleic acid sequence"/"given viral nucleic acid sequence" x 100. We estimated the yield for both sequences as 59%. Moreover, we studied the influence of human serum, dsDNA and non-complementary sequence of nucleic acids on isolation of viral nucleic acids. We also used carbon nanotubes-based screen-printed electrodes coupled with micro-flow instrument to detect viral nucleic acids. We were able to isolate and detect nanogram amounts of nucleic acids.
Keywords
Paramagnetic microparticles; Viral nucleic acid; Electrochemical detection; Voltammetry; Screen-printed electrode
Authors
ADAM, V.; HÚSKA, D.; HUBÁLEK, J.; KIZEK, R.
RIV year
2010
Released
1. 3. 2010
Publisher
SPRINGER HEIDELBERG
Location
TIERGARTENSTRASSE 17, D-69121 HEIDELBERG, GERMANY
ISBN
1613-4982
Periodical
Microfluidics and Nanofluidics
Year of study
8
Number
3
State
Federal Republic of Germany
Pages from
329
Pages to
339
Pages count
11
BibTex
@article{BUT50229, author="Vojtěch {Adam} and Dalibor {Húska} and Jaromír {Hubálek} and René {Kizek}", title="Easy to use and rapid isolation and detection of a viral nucleic acid by using paramagnetic microparticles and carbon nanotubes-based screen-printed electrodes", journal="Microfluidics and Nanofluidics", year="2010", volume="8", number="3", pages="329--339", issn="1613-4982" }