title: Fluorescent immunochromatography for rapid and sensitive typing of seasonal influenza viruses.
abstract: Lateral flow tests also known as Immunochromatography (IC) is an antigen-detection method conducted on a nitrocellulose membrane that can be completed in less than 20 min. IC has been used as an important rapid test for clinical diagnosis and surveillance of influenza viruses, but the IC sensitivity is relatively low (approximately 60%) and the limit of detection (LOD) is as low as 10³ pfu per reaction. Recently, we reported an improved IC assay using antibodies conjugated with fluorescent beads (fluorescent immunochromatography; FLIC) for subtyping H5 influenza viruses (FLIC-H5). Although the FLIC strip must be scanned using a fluorescent reader, the sensitivity (LOD) is significantly improved over that of conventional IC methods. In addition, the antibodies which are specific against the subtypes of influenza viruses cannot be available for the detection of other subtypes when the major antigenicity will be changed. In this study, we established the use of FLIC to type seasonal influenza A and B viruses (FLIC-AB). This method has improved sensitivity to 100-fold higher than that of conventional IC methods when we used several strains of influenza viruses. In addition, FLIC-AB demonstrated the ability to detect influenza type A and influenza type B viruses from clinical samples with high sensitivity and specificity (Type A: sensitivity 98.7% (74/75), specificity 100% (54/54), Type B: sensitivity 100% (90/90), specificity 98.2% (54/55) in nasal swab samples) in comparison to the results of qRT-PCR. And furthermore, FLIC-AB performs better in the detection of early stage infection (under 13 h) than other conventional IC methods. Our results provide new strategies to prevent the early-stage transmission of influenza viruses in humans during both seasonal outbreaks and pandemics.
authors: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.
authors: Akira
authors: A
authors: Sakurai
authors: ADTEC Co., LTD., 1693-6 Yokkaichi, Usa-City, Oita, 879-0471, Japan.
authors: Katsuyoshi
authors: K
authors: Takayama
authors: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.
authors: Namiko
authors: N
authors: Nomura
authors: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.
authors: Naoki
authors: N
authors: Kajiwara
authors: Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, 060-0818, Japan.
authors: Masatoshi
authors: M
authors: Okamatsu
authors: Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, 060-0818, Japan; Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, 2-1-6, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.
authors: Naoki
authors: N
authors: Yamamoto
authors: Konica Minolta, Inc., No.1 sakura-machi, Hino-shi, Tokyo, 191-8511, Japan.
authors: Tsuruki
authors: T
authors: Tamura
authors: Konica Minolta, Inc., No.1 sakura-machi, Hino-shi, Tokyo, 191-8511, Japan.
authors: Jitsuho
authors: J
authors: Yamada
authors: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.
authors: Masako
authors: M
authors: Hashimoto
authors: Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, 060-0818, Japan; Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo, 001-0020, Japan.
authors: Yoshihiro
authors: Y
authors: Sakoda
authors: Konica Minolta, Inc., No.1 sakura-machi, Hino-shi, Tokyo, 191-8511, Japan.
authors: Yoshihiko
authors: Y
authors: Suda
authors: ADTEC Co., LTD., 1693-6 Yokkaichi, Usa-City, Oita, 879-0471, Japan.
authors: Yukuharu
authors: Y
authors: Kobayashi
authors: Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, 060-0818, Japan; Global Station for Zoonosis Control, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo, 001-0020, Japan.
authors: Hiroshi
authors: H
authors: Kida
authors: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.
authors: Futoshi
authors: F
authors: Shibasaki
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D001717:Birds; D002648:Child; D002675:Child, Preschool; D002846:Chromatography, Affinity / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D009980:Influenza A virus / Q000145:classification* / Q000302:isolation & purification; D009981:Influenza B virus / Q000145:classification* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis*; D008297:Male; D008322:Mammals / Q000821:virology; D008875:Middle Aged; D058889:Molecular Typing / Q000379:methods*; D009976:Orthomyxoviridae Infections / Q000175:diagnosis; D012680:Sensitivity and Specificity; D055815:Young Adult
references: N Engl J Med. 2013 May 16;368(20):1888-97
references: 23577628
references: CMAJ. 2010 Feb 9;182(2):131-6
references: 19959592
references: Lancet. 2013 Jun 1;381(9881):1916-25
references: 23623390
references: Lancet. 1998 Feb 14;351(9101):472-7
references: 9482438
references: Nature. 2013 Sep 26;501(7468):556-9
references: 23842497
references: Eur J Pediatr. 2011 Apr;170(4):511-7
references: 20938682
references: Science. 2009 Jun 19;324(5934):1557-61
references: 19433588
references: Euro Surveill. 2013 Apr 11;18(15):20453
references: 23594575
references: Lancet. 2013 Jun 1;381(9881):1926-32
references: 23643111
references: Virology. 2012 Jan 5;422(1):151-60
references: 22078166
references: Virology. 1990 Mar;175(1):59-68
references: 2309452
references: Viruses. 2012 Aug;4(8):1235-57
references: 23012622
references: mBio. 2013 Jul 02;4(4):
references: 23820393
references: N Engl J Med. 2006 Nov 23;355(21):2174-7
references: 17124014
references: N Engl J Med. 2009 Jun 18;360(25):2605-15
references: 19423869
references: Virology. 1988 Mar;163(1):112-22
references: 3267218
references: PLoS Curr. 2009 Dec 20;1:RRN1139
references: 20043033
references: Nature. 2013 Sep 26;501(7468):551-5
references: 23842494
references: PLoS Med. 2009 Dec;6(12):e1000207
references: 19997612
references: J Virol. 1990 Jun;64(6):2860-5
references: 2335820
references: Arch Virol Suppl. 1997;13:105-13
references: 9413531
references: PLoS One. 2012;7(11):e50670
references: 23226350
references: Science. 1998 Jan 16;279(5349):393-6
references: 9430591
references: Ann Emerg Med. 2006 Mar;47(3):250-4
references: 16492491
references: J Virol Methods. 2011 Dec;178(1-2):75-81
references: 21889540
references: N Engl J Med. 2009 Nov 12;361(20):1935-44
references: 19815859
references: BMJ. 2009 Jul 14;339:b2840
references: 19602714
references: Nature. 1997 Oct 9;389(6651):554
references: 9335492
references: J Clin Virol. 2007 Feb;38(2):169-71
references: 17194622
references: Emerg Microbes Infect. 2012 Oct;1(10):e33
references: 26038404
references: MMWR Morb Mortal Wkly Rep. 2012 Sep 28;61:764-7
references: 23013722
references: Lancet. 2008 Apr 26;371(9622):1464-75
references: 18440429
references: J Virol Methods. 2011 Dec;178(1-2):82-6
references: 21911008
references: Vet Microbiol. 2013 May 31;164(1-2):39-45
references: 23462521
references: J Clin Microbiol. 2010 Aug;48(8):2872-7
references: 20554831
references: PLoS One. 2013 Nov 06;8(11):e76753
references: 24223117