title: Influenza virus detection with pentabody-activated nanoparticles.
abstract: A nanoparticle-based immunoassay was developed for the rapid and sensitive detection of avian influenza virus (AIV). In this method, AIV-specific pentabody (pVHH3B) was conjugated to magnetic nanoparticles (MNPs) and used to capture AIV. Gold nanoparticles (GNPs), labelled with the anti-AIV mouse monoclonal antibody 3C8, were used as a detector. In the presence of target samples, the pentabody pVHH3B enriched AIV on the MNPs. Thereafter, mAb 3C8-labelled GNPs (GNPs-mAb3C8) bound to MNPs via AIV and were separated using a magnetic field. GNPs in the complex catalyzed the oxidation of hydroquinone to quinone, and the OD value of quinone was measured. The developed assay displayed substantial signal change after incubation in an AIV sample in a concentration-dependent manner. The detection limit was 10 ng/ml, which is 10 times more sensitive than conventional double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). In conclusion, by combining MNPs and a novel pentabody pVHH3B, this study provided a sensitive influenza viral detection assay that has the potential to become a rapid, sensitive and inexpensive diagnostic tool for infectious diseases.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, 100101 Beijing, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Mu
authors.forename: Xinglu
authors.initials: X
authors.lastname: Huang
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Bu
authors.forename: Jie
authors.initials: J
authors.lastname: Zhuang
authors.forename: Zhixue
authors.initials: Z
authors.lastname: Cheng
authors.forename: Jing
authors.initials: J
authors.lastname: Feng
authors.forename: Dongling
authors.initials: D
authors.lastname: Yang
authors.forename: Changsheng
authors.initials: C
authors.lastname: Dong
authors.forename: Jianbing
authors.initials: J
authors.lastname: Zhang
authors.forename: Xiyun
authors.initials: X
authors.lastname: Yan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D016227:Benzoquinones / Q000378:metabolism; D001717:Birds; D006801:Humans; D006873:Hydroquinones / Q000378:metabolism; D007118:Immunoassay / Q000379:methods; D018189:Immunomagnetic Separation / Q000379:methods*; D009980:Influenza A virus / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D051379:Mice; D053758:Nanoparticles; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D013053:Spectrophotometry / Q000379:methods; D014773:Virology / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D016227:Benzoquinones; D006873:Hydroquinones; C004532:quinone; C031927:hydroquinone