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Title: MULTIPLEXED IMMUNOASSAYS FOR VIRAL PATHOGENS UTILIZING SURFACE-ENHANCED RAMAN SCATTERING

Author
item DRISKELL, JEREMY - IOWA STATE UNIVERSITY
item KWARTA, KAREN - IOWA STATE UNIVERSITY
item LIPERT, ROBERT - IOWA STATE UNIVERSITY
item Neill, John
item Ridpath, Julia
item PORTER, MARC - IOWA STATE UNIVERSITY

Submitted to: Proceedings of the Office International Des Epizooties World Association of
Publication Type: Abstract Only
Publication Acceptance Date: 7/15/2005
Publication Date: 10/3/2006
Citation: Driskell, J., Kwarta, K., Lipert, R.J., Neill, J.D., Ridpath, J.F., Porter, M.D. 2006. Multiplexed immunoassays for viral pathogens utilizing surface-enhanced raman scattering [abstract]. New Diagnostic Technology: Applications in Animal Health & Biologics Controls International Congress. Office International Des Epizooties. p. 70.

Interpretive Summary:

Technical Abstract: A sandwich immunoassay for the selective detection of viral pathogens has been developed using surface-enhanced Raman scattering (SERS) as the readout technology. The strengths of SERS-based detection include ultrahigh sensitivity and the possibility of multiplexing using multiple tags. The viability of this approach is demonstrated by the binding of feline calicivirus (FCV) from cell culture lysate to a capture substrate that is coated with anti-FCV monoclonal antibodies (mAbs). To detect binding, the surface-bound FCV is tagged with an extrinsic Raman label (ERL) which allows identification and quantification. ERLs consist of 60-nm gold nanoparticles (which provide a large enhancement factor), a Raman reporter molecule coating (which engenders a unique spectral signature), and a mAb coating (which enables bioselectivity). This presentation outlines the development and optimization of a chip-scale assay that allows visualization and quantitation of individual binding events. Efforts to minimize nonspecific binding, maximize FCV binding efficiency, and improve the detection limit are discussed. Results that apply this approach to the detection of parvovirus in a multiplexed assay are also briefly described.