Development of an antigen-capture enzyme-linked immunosorbent assay using monoclonal antibodies for detecting H6 avian influenza viruses

Abstract

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 antigen-capture ELISA could be used to specifically detect H6N1 AIV.

Optical fiber sensor based on surface plasmon resonance for rapid detection of avian influenza virus subtype H6: Initial studies

“…Thus, to control AIVs before they spread, a method for detecting viruses or antigens at an early stage must be applied. Chen et al (2012) developed an AC-ELISA for detecting AIV-H6.…”

“…Tracheal samples from market-age Taiwan Country chickens (10-15 weeks old) were prepared as described by Chen et al, 2012. In brief, the mucosa was scraped from each of the tracheal samples and homogenized with 1 mL of tryptose phosphate.…”

Development of a monoclonal antibody‐based antigen capture enzyme‐linked immunosorbent assay for detection of H7N9 subtype avian influenza virus

“…The binding affinity of mAbs was evaluated by indirect ELISA as described previously. 7 The specificity was detected by immunofluorescence assay (IFA) as described previously. 11 The control mAb (1E7) against syphilis and phosphate-buffered saline (PBS) were used as negative controls.…”

Immuno Nanosensor for the Ultrasensitive Naked Eye Detection of Tuberculosis

“…However, these conventional techniques are laborious and time consuming due to the slow-growing type M. tuberculosis . In order to improve TB detection, various biosensing techniques such as nucleic acid amplification tests [ 7 , 8 , 9 ], latex agglutination [ 10 ], enzyme-linked immunosorbent assay (ELISA) [ 11 ], radiometric detection [ 12 ], and flow cytometry [ 13 ] have been implemented. These techniques provide rapid and improved sensitivity and specificity functions for diagnosis of TB but many have limitations with regard to providing real-time detection results and also require highly skilled personnel to handle the lengthy procedures and complex instrumentation.…”

“…Other tests such as ELISA, flow cytometry, etc. are available, but they too suffer from limitations associated with high costs, complex assay handling, the need for expensive instruments and trained personnel, and the difficulties of transferring the test to remote rural regions, especially in developing nations [ 11 , 12 , 13 ]. In order to efficiently diagnose TB, the test should be widely available, user-friendly, require minimal or no-training of personnel and produce highly accurate outcomes which are available for use in areas with peripheral resources.…”