|Bowen, J -|
|Mecham, J -|
|Hamlin, M -|
|Henderson, B -|
|Kim, M -|
|Mirjankar, N -|
|Lavine, B -|
Submitted to: Microchemical Journal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: June 14, 2011
Publication Date: June 21, 2011
Repository URL: http://ac.els-cdn.com/S0026265X11001275/1-s2.0-S0026265X11001275-main.pdf?_tid=89019f6d9ca9b1035eb8caad0d0f4b8e&acdnat=1345831397_f897a36c605f921ce6b3beaff0040fe1
Citation: Bowen, J., Mecham, J.0., Hamlin, M., Henderson, B., Kim, M., Mirjankar, N., Lavine, B.K. 2011. Development of field-deployable instrumentation based on “antigen–antibody” reactions for detection of hemorrhagic disease in ruminants. Microchemical Journal. Interpretive Summary: Rapid diagnostic tests to detect blue tongue virus (BTV) and epizootic hemorrhage disease virus (EHDV) in wild and domestic animals is critical for identification and control of disease outbreaks. A biosensor was designed to detect virus antigen and antibody to virus from animal samples in 20 min. The antigen biosensor was able to detect and differentiate between the two closely related viruses. The antibody biosensor was able to detect antibody but could not differentiate between the two viruses.
Technical Abstract: Development of field-deployable methodology utilizing antigen–antibody reactions and the surface Plasmon resonance (SPR) effect to provide a rapid diagnostic test for recognition of the blue tongue virus (BTV) and epizootic hemorrhage disease virus (EHDV) in wild and domestic ruminants is reported. A Spreeta chip, which utilizes microelectronic technology to implement the SPR effect, is shown to possess sufficient sensitivity and operating speed to detect either BTV and EHVD antigens or antibodies in real time. The biosensor consists of an outer active surface layer comprised of either an antibody or antigen immobilized by covalent bonds through several other organic layers including a self assembled monolayer to a gold surface. Parallel experiments were run on the biosensor surface using either a home-built high resolution SPR instrument or a low resolution solid state Spreeta SPR chip. Both instruments were capable of monitoring the antigen–antibody reaction used to selectively detect the presence of BTV and EHDV viral pathogens. Results for the antibody and antigen reactive layers with antigen or antibody solutions as well as the modeling of these layers are discussed. The characteristics of these biosensors – specificity and time of reaction – were assessed. The antibody surface biosensors exhibited a high degree of specificity, even when using low resolution instrumentation. The time of analysis was under 20 min, which was the arbitrary exposure time.