Integrated microdevice of reverse transcription-polymerase chain reaction with colorimetric immunochromatographic detection for rapid gene expression analysis of influenza A H1N1 virus

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An integrated microdevice of a reverse transcription-polymerase chain reaction (RT-PCR) reactor and an immunochromatographic strip was constructed for colorimetric detection of gene expression of influenza A virus subtype H1N1. An RT-PCR cocktail, which included Texas Red-labeled primers, dNTP including biotin-labeled dUTP, and RNA templates of influenza A H1N1 virus, was filled in the PCR chamber through the micropump, and the RT-PCR was performed to amplify the target H1 gene (102 bp). The resultant amplicons bearing biotin moieties and Texas Red haptens were subsequently eluted to the immunochromatographic strip, in which they were first conjugated with the gold nanoparticle labeled anti-hapten antibody in the conjugation pad, and then captured on the streptavidin coated test line through the biotin-streptavidin interaction. By observing a violet color in the test line which was derived from the gold nanoparticle, we confirmed the HI NI target virus. The entire process on the integrated microdevice consisting of a micropump, a 2 mu L PCR chamber, and an immunochromatographic strip was carried out on the portable genetic analyzer within 2.5 h, enabling on-site colorimetric pathogen identification with detection sensitivity of 14.1 pg RNA templates. (C) 2011 Elsevier B.V. All rights reserved.
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Issue Date
2012-03
Language
English
Article Type
Article
Keywords

CAPILLARY ELECTROPHORETIC MICRODEVICE; FUNCTIONALIZED GOLD NANOPARTICLES; REAGENT STRIP BIOSENSOR; MICROFLUIDIC CASSETTE; PATHOGEN DETECTION; DNA; MICROSYSTEM; AMPLIFICATION; PROBES; SARS

Citation

BIOSENSORS & BIOELECTRONICS, v.33, no.1, pp.88 - 94

ISSN
0956-5663
DOI
10.1016/j.bios.2011.12.024
URI
http://hdl.handle.net/10203/98100
Appears in Collection
CBE-Journal Papers(저널논문)
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