title: A packaged paper fluidic-based microdevice for detecting gene expression of influenza A virus.
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 10(2) copies of RNA templates in 10 min. Pathotyping and subtyping of the clinical nasopharyngeal swab samples were also analyzed whose results were confirmed by real-time RT-PCR.
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Yong Tae
authors.initials: YT
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jae Hwan
authors.initials: JH
authors.lastname: Jung
authors.affiliation: Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University, 12 Gaeshin-Dong, Heungduk-gu, Cheongju-si, Chungcheongbuk-do 361-763, Republic of Korea.
authors.forename: Young Ki
authors.initials: YK
authors.lastname: Choi
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: seots@kaist.ac.kr.
authors.forename: Tae Seok
authors.initials: TS
authors.lastname: Seo
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation; D004867:Equipment Design; D015870:Gene Expression; D060005:Genotyping Techniques / Q000295:instrumentation*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000302:isolation & purification; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics* / Q000302:isolation & purification; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D057230:Limit of Detection; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D060885:Multiplex Polymerase Chain Reaction / Q000295:instrumentation; D010209:Paper; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D011934:Reagent Strips / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation
chemical_list: D012367:RNA, Viral; D011934:Reagent Strips
keywords: Gene expression; Immunochromatographic strip; Influenza A virus; Reverse transcriptase-polymerase chain reaction; Subtyping