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Volume 45, Number 3, June 2012

Development of an antigen-capture enzyme-linked immunosorbent assay using monoclonal antibodies

for detecting H6 avian influenza viruses

Yi-Tung Chen, Zak Tsao, Shu-Ting Chang, Ron-Huay Juang, Lih-Chiann Wang, Chung-Ming Chang, Ching-Ho Wang

Received: January 4, 2011    Revised: July 12, 2011    Accepted: August 1, 2011   


Corresponding author:

 School of Veterinary Medicine, National Taiwan University, No. 1, Sec. 4, Roosevelt Road, Taipei 10617,

Taiwan. E-mail address: (C.-H. Wang).


Background and purpose: 

 The H6 subtype of avian influenza virus (AIV) infection occurs frequently in wild and domestic birds. AIV antigen detection is preferred for controlling AIV as birds are infected before they produce antibodies. The purpose of this study was to develop an early diagnostic method for AIV detection. Six monoclonal antibodies (mAbs) developed from a field H6N1 AIV strain were tested for their ability to bind to viruses. The two that showed the greatest binding ability to AIVs were used for antigen detection. An antigen-capture enzyme-linked immunosorbent assay (ELISA) to detect H6 AIVs was developed using these mAbs. One mAb was coated onto an ELISA plate as the capture antibody. The other mAb was used as the detector antibody after labeling with horseradish peroxidase. The antigen-capture ELISA detected

H6N1 AIVs but not H5 AIVs, human H1N1, H3N2 influenza or other viruses. This antigencapture ELISA could be used to specifically detect H6N1 AIV.


Key words:

 Antigen-capture enzyme-linked immunosorbent assay (AC-ELISA); Avian influenza virus; H6