A packaged paper fluidic-based microdevice for detecting gene expression of influenza A virus

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Pathotyping and subtyping of influenza A virus were performed with a packaged paper fluidic-based analytical microdevice (PFAM) after one-step reverse transcription-polymerase chain reaction (RT-PCR). The PFAM contains two test lines: one for detecting M gene to identify the influenza A virus and another for haemagglutinin subtyping to determine the viral strain among H1N1, H3N2, and H5N1. The M gene and the haemagglutinin gene (H1, H3, and H5 genes) were amplified by using the Digoxigenin and the Texas Red modified primers, respectively, in the multiplex RT-PCR. The amplicon products were loaded in the conjugate pad of the PFAM in which the streptavidin coated gold nanoparticles were linked with the biotin moieties that were incorporated in the middle of the DNA strands, and then captured by the anti-Digoxigenin and anti-Texas Red immobilized on the test lines. Influenza A H1N1, H3N2, and H5N1 could be identified with a limit of detection of 10(2) copies of RNA templates in 10 mm. Pathotyping and subtyping of the clinical nasopharyngeal swab samples were also analyzed whose results were confirmed by real-time RT-PCR.
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Issue Date
2014-11
Language
English
Article Type
Article
Keywords

AVIAN INFLUENZA; RAPID DETECTION; H5N1 VIRUS; IMMUNOCHROMATOGRAPHIC STRIP; ASSAY; CULTURE; NASBA; PCR; EMERGENCE; BINDING

Citation

BIOSENSORS BIOELECTRONICS, v.61, pp.485 - 490

ISSN
0956-5663
DOI
10.1016/j.bios.2014.06.006
URI
http://hdl.handle.net/10203/192353
Appears in Collection
CBE-Journal Papers(저널논문)
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