Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test

To monitor the progression of infectious diseases, it is useful to assess immunoreactivity against various antigenic determinants, and measure different antibody isotypes because they appear at different stages of the host immune response. With Lyme borreliosis, the pathogenic agent can be one of the multiple members of the Borrelia species. Therefore, correct sample classification requires evaluating the immunoreactivity against different antigens of different Borrelia species. Additionally, anti-pathogen IgG and IgM responses can have different elicitation time courses during disease progression. Here we demonstrate the development of a two -reporter multiplex immunoassay that has utility in identifying Borrelia-specific immune response in human serum samples by simultaneously evaluating both IgG and IgM immunoreactivity against different bacterial antigens in the same reaction well. This dual-reporter approach retains the analytical performance of single-reporter methods while conserving time and resources and reducing sample size requirements. This assay allows essentially double the serological information to be generated from a blood sample in half the time.

Automated summary

To monitor infectious diseases, it is important to assess immunoreactivity and measure different antibody isotypes at different stages of the immune response. Lyme borreliosis can be caused by various Borrelia species, so correct classification of samples requires evaluating immunoreactivity against different antigens. This study demonstrates a dual-reporter multiplex immunoassay that allows simultaneous evaluation of IgG and IgM immunoreactivity against different bacterial antigens, providing more serological information in less time.