Integrated microdevice of reverse transcription-polymerase chain reaction with colorimetric immunochromatographic detection for rapid gene expression analysis of influenza A H1N1 virus.
作者： Yong Tae Kim , Yuchao Chen , Jong Young Choi , Won-Jung Kim , Hyun-Mi Dae
DOI: 10.1016/J.BIOS.2011.12.024
关键词:
摘要: Abstract An integrated microdevice of a reverse transcription-polymerase chain reaction (RT-PCR) reactor and an immunochromatographic strip was constructed for colorimetric detection gene expression influenza A virus subtype H1N1. RT-PCR cocktail, which included Texas Red-labeled primers, dNTP including biotin-labeled dUTP, RNA templates H1N1 virus, filled in the PCR chamber through micropump, performed to amplify target H1 (102 bp). The resultant amplicons bearing biotin moieties Red haptens were subsequently eluted strip, they first conjugated with gold nanoparticle labeled anti-hapten antibody conjugation pad, then captured on streptavidin coated test line biotin–streptavidin interaction. By observing violet color derived from nanoparticle, we confirmed virus. entire process consisting 2 μL chamber, carried out portable genetic analyzer within 2.5 h, enabling on-site pathogen identification sensitivity 14.1 pg templates.
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