Development of a whole-virus multiplex flow cytometric assay for antibody screening of a specific pathogen-free primate colony.
Our goal was to determine if a multiplex technique using a fluorescent bead-based flow cytometric assay could yield results comparable to traditional enzyme-linked immunosorbent assay (ELISA) in terms of sensitivity, specificity, cross-reactivity, and throughput. We applied both techniques to serologic screening of specific pathogen-free macaques, for type D simian retrovirus, simian T-lymphotropic virus, Cercopithicine herpesvirus 1, and simian immunodeficiency virus, and found a high correlation between the bead-based multiplex assay and ELISA. The multiplex assay demonstrated greater sensitivity with no loss in specificity when compared to the ELISA. A lower false-positive rate with the multiplex assay decreased the number of confirmatory Western blots required. Using the multiplex assay, we were able to screen samples for 4 viruses simultaneously in the time it took to perform a single-virus ELISA, resulting in a faster turnaround time and higher throughput. The multiplexed assay provided greater sensitivity, increased stability, and better performance than ELISA.
Kuller
LaRene
L
Washington National Primate Research Center, University of Washington, Box 357331, Seattle, 98195, USA.
Watanabe
Robin
R
Anderson
David
D
Grant
Richard
R
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006568:Herpesvirus 1, Cercopithecine / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood; D008863:Microspheres; D015301:Retroviruses, Simian / Q000276:immunology; D012680:Sensitivity and Specificity; D015302:Simian Immunodeficiency Virus / Q000276:immunology; D015303:Simian T-lymphotropic virus 1 / Q000276:immunology; D013047:Specific Pathogen-Free Organisms; D014777:Virus Diseases / Q000175:diagnosis* / Q000276:immunology* / Q000821:virology; D014780:Viruses / Q000276:immunology*
D000914:Antibodies, Viral; D007074:Immunoglobulin G