This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ho, H.-T.
Right arrow Articles by Kwang, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ho, H.-T.
Right arrow Articles by Kwang, J.

 Previous Article  |  Next Article 

Clinical and Vaccine Immunology, May 2009, p. 726-732, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00465-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Rapid Detection of H5N1 Subtype Influenza Viruses by Antigen Capture Enzyme-Linked Immunosorbent Assay Using H5- and N1-Specific Monoclonal Antibodies {triangledown}

Hui-Ting Ho,1 Hong-Liang Qian,1 Fang He,1 Tao Meng,1 Milene Szyporta,1 Nayana Prabhu,1 Mookkan Prabakaran,1 Kwai-Peng Chan,2 and Jimmy Kwang1,3*

Animal Health Biotechnology, Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604, Singapore,1 Department of Pathology, Singapore General Hospital, Singapore 169608, Singapore,2 Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore, Singapore3

Received 10 December 2008/ Returned for modification 10 February 2009/ Accepted 13 March 2009

Highly pathogenic avian influenza (HPAI) virus of the H5N1 subtype has caused devastating damage to poultry flocks and sporadic human H5N1 infections. There is concern that this virus subtype may gain transmissibility and become pandemic. Rapid diagnosis and surveillance for H5N1 subtype viruses are critical for the control of H5N1 infection. In this study, we report a robust antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) based on H5- and N1-specific monoclonal antibodies (MAbs) for the rapid detection of H5N1 subtype viruses. The H5 hemagglutinin (HA)-specific MAb (2D9) targets a conformational epitope which recognized multiple clades of H5N1 viruses, including clades 0, 1, 2.1, 2.2, 2.3, 4, 7, and 8. The N1 neuraminidase (NA)-specific MAb (8H12) recognized a linear epitope comprising the sequence AELPF. This epitope was 99% conserved in the NA of 708 analyzed H5N1 viruses, while the epitope was absent in NAs of subtypes N2 through N9. The specificity of the AC-ELISA was examined by using 41 H5N1 HPAI strains from multiple clades, 36 non-H5N1 viruses, and 4 influenza B viruses. No cross-reactivity was observed for any of the non-H5N1 viruses tested. The estimated detection limit was 1 to 2 HA titers. It is concluded that this H5N1 AC-ELISA can simultaneously detect H5 and N1 subtype antigens, eliminating the need for secondary testing for the NA subtype. Implementation of this assay in ELISA-like formats suitable for field use, such as dot ELISA, immunofiltration, or electrochemical biosensor technologies, would provide dual on-site detection of H5 and N1 in clinical or environmental specimens.

* Corresponding author. Mailing address: Animal Health Biotechnology, Temasek Life Science Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore. Phone: 65-68727473. Fax: 65-68727007. E-mail: kwang{at}tll.org.sg

{triangledown} Published ahead of print on 25 March 2009.

Clinical and Vaccine Immunology, May 2009, p. 726-732, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00465-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»