An RNA aptamer that distinguishes between closely related human influenza viruses and inhibits haemagglutinin-mediated membrane fusion

• Published in: Journal of general virology 2006-03, Vol.87 (3), p.479-487
• Main Authors: Gopinath, Subash C. B, Misono, Tomoko S, Kawasaki, Kazunori, Mizuno, Takafumi, Imai, Masaki, Odagiri, Takato, Kumar, Penmetcha K. R
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Aptamers, Nucleotide - genetics; Aptamers, Nucleotide - metabolism; Base Sequence; Binding Sites - genetics; Biological and medical sciences; Erythrocyte Membrane - physiology; Erythrocyte Membrane - virology; Fundamental and applied biological sciences. Psychology; Hemagglutinin Glycoproteins, Influenza Virus - chemistry; Hemagglutinin Glycoproteins, Influenza Virus - metabolism; Hemagglutinin Glycoproteins, Influenza Virus - physiology; Humans; Influenza A virus; Influenza A Virus, H3N2 Subtype - classification; Influenza A Virus, H3N2 Subtype - metabolism; Membrane Fusion; Microbiology; Miscellaneous; Models, Molecular; Molecular Sequence Data; Nucleic Acid Conformation; Protein Binding; SELEX Aptamer Technique; Sensitivity and Specificity; Sequence Alignment; Species Specificity; Virology
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/vir.0.81508-0

1 Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Central 6, 1-1 Higashi, Tsukuba City, Ibaraki 305-8566, Japan 2 Department of Virology III, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan Correspondence Penmetcha K. R. Kumar pkr-kumar{at}aist.go.jp Aptamers selected against various kinds of targets have shown remarkable specificity and affinity, similar to those displayed by antibodies to their antigens. To employ aptamers as genotyping reagents for the identification of pathogens and their strains, in vitro selections were carried out to find aptamers that specifically bind and distinguish the closely related human influenza A virus subtype H3N2. The selected aptamer, P30-10-16, binds specifically to the haemagglutinin (HA) region of the target strain A/Panama/2007/1999(H3N2) and failed to recognize other human influenza viruses, including another strain with the same subtype, H3N2. The aptamer displayed over 15-fold-higher affinity to the HA compared with the monoclonal antibody, and efficiently inhibited HA-mediated membrane fusion. These studies delineate the application of aptamers in the genotyping of viruses. Published online ahead of print on 29 November 2005 as DOI 10.1099/vir.0.81508-0. Supplementary methods, figures and videos are available in JGV Online. These authors contributed equally to this work.