Broad-spectrum detection of H5 subtype influenza A viruses with a new fluorescent immunochromatography system.
Immunochromatography (IC) is an antigen-detection assay that plays an important role in the rapid diagnosis of influenza virus because the protocol is short time and easy to use. Despite the usability of IC, the sensitivity is approximately 10(3) pfu per reaction. In addition, antigen-antibody interaction-based method cannot be used for the detection of influenza viruses with major antigenic change. In this study, we established the use of fluorescent immunochromatography (FLIC) to detect a broad spectrum of H5 subtype influenza A viruses. This method has improved sensitivity 10-100 fold higher than traditional IC because of the use of fluorescent conjugated beads. Our Type-E FLIC kit detected all of the H5 subtype influenza viruses that were examined, as well as recombinant hemagglutinin (HA) proteins (rHAs) belonging to the Eurasian H5 subtype viruses and the Type-N diagnosed North American H5 subtype influenza A viruses. Thus, this kit has the improved potential to detect H5 subtype influenza viruses of different clades with both Type-E and Type-N FLIC kits. Compared with PCR-based diagnosis, FLIC has a strong advantage in usability, because the sample preparation required for FLIC is only mix-and-drop without any additional steps such as RNA extraction. Our results can provide new strategies against the spread and transmission of HPAI H5N1 viruses in birds and mammals including humans.
Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya-ku, Tokyo, Japan.
Akira
A
Sakurai
Katsuyoshi
K
Takayama
Namiko
N
Nomura
Tsubasa
T
Munakata
Naoki
N
Yamamoto
Tsuruki
T
Tamura
Jitsuho
J
Yamada
Masako
M
Hashimoto
Kazuhiko
K
Kuwahara
Yoshihiro
Y
Sakoda
Yoshihiko
Y
Suda
Yukuharu
Y
Kobayashi
Nobuo
N
Sakaguchi
Hiroshi
H
Kida
Michinori
M
Kohara
Futoshi
F
Shibasaki
D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry; D000914:Antibodies, Viral / Q000737:chemistry; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D004285:Dogs; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D057230:Limit of Detection; D061985:Madin Darby Canine Kidney Cells; D011933:Reagent Kits, Diagnostic; D013050:Spectrometry, Fluorescence
D055910:Antibodies, Immobilized; D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011933:Reagent Kits, Diagnostic; C483468:hemagglutinin, human influenza A virus