tetano
Editor, Senior Moderator
Biosens Bioelectron. 2014 Jun 8;61C:485-490. doi: 10.1016/j.bios.2014.06.006. [Epub ahead of print]
A packaged paper fluidic-based microdevice for detecting gene expression of influenza A virus.
Kim YT1, Jung JH1, Choi YK2, Seo TS3.
Author information
Abstract
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 102 copies of RNA templates in 10min. Pathotyping and subtyping of the clinical nasopharyngeal swab samples were also analyzed whose results were confirmed by real-time RT-PCR.
Copyright ? 2014 Elsevier B.V. All rights reserved.
KEYWORDS:
Gene expression; Immunochromatographic strip; Influenza A virus; Reverse transcriptase-polymerase chain reaction; Subtyping
PMID:
24949821
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24949821
A packaged paper fluidic-based microdevice for detecting gene expression of influenza A virus.
Kim YT1, Jung JH1, Choi YK2, Seo TS3.
Author information
Abstract
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 102 copies of RNA templates in 10min. Pathotyping and subtyping of the clinical nasopharyngeal swab samples were also analyzed whose results were confirmed by real-time RT-PCR.
Copyright ? 2014 Elsevier B.V. All rights reserved.
KEYWORDS:
Gene expression; Immunochromatographic strip; Influenza A virus; Reverse transcriptase-polymerase chain reaction; Subtyping
PMID:
24949821
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24949821