The development of vaccines against malaria and serodiagnostic tests for detecting recent exposure requires tools for antigen discovery and suitable animal models. The protein microarray is a high throughput, sample sparing technique, with applications in infectious disease research, clinical diagnostics, epidemiology, and vaccine development. We recently demonstrated Qdot®-based indirect immunofluorescence together with a portable optical imager ArrayCAM® using single isotype detection could replicate data using the conventional laser confocal scanner system, opening up the opportunity for deployment of protein microarray-based immunoassays in more laboratories worldwide. We developed a multiplexing protocol for simultaneous detection of IgG, IgA and IgM and compared samples from a controlled human malaria infection model with those from controlled malaria infections of Aotus nancymaae , a widely-used non-human primate model of human malaria. IgG profiles showed the highest concordance in number of reactive antigens. Thus, of the 139 antigens recognized by human IgG antibody, 111 were also recognized by Aotus monkeys. Interestingly, IgA profiles were largely non-overlapping. Finally, on the path toward wider deployment of the portable platform, we show excellent correlations between array data obtained in 5 independent laboratories around the U.S. using the multiplexing protocol ( R 2 : 0.60 – 0.92).This study supports the use of this platform for wider deployment, particularly in endemic areas where such a tool will have the greatest impact on global human health.
【저자키워드】 diagnostics, immunoassay, protein microarray, multiplex, antibody isotype, quantum dots,