Rapid detection of anti-H5 avian influenza virus antibody by fluorescence polarization immunoassay using a portable fluorescence polarization analyzer

利用便携式荧光偏振分析仪通过荧光偏振免疫测定法快速检测抗H5禽流感病毒抗体

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作者:Keine Nishiyama,Yohei Takeda,Masatoshi Maeki,Akihiko Ishida,Hirofumi Tani,Koji Shigemura,Akihide Hibara,Yutaka Yonezawa,Kunitoshi Imai,Haruko Ogawa,Manabu Tokeshi

Abstract

A rapid, facile and selective detection of anti-H5 subtype avian influenza virus (AIV) antibody in serum by fluorescence polarization immunoassay (FPIA) was achieved. A fragment of recombinant H5 subtype AIV hemagglutinin was produced and labeled with fluorescein to use it as a labeled antigen in FPIA. This labeled antigen was mixed with anti-AIV sera (H1-H16 subtypes) and FP of the mixture was measured using a portable FP analyzer on a microdevice. It was found that FP increased in proportion to the concentration of anti-H5 AIV antibody (serum) and was significantly higher than FP obtained with the other sera. The selective detection of anti-H5 subtype AIV antibody was confirmed. The required volume of original sample was 2 μL and analysis time was within 20 min. This detection system realizes an efficient on-site diagnosis and surveillance of AIV.

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