Lyme disease, transmitted to humans through the bite of the black-legged tick, is the most common vector-borne disease in the United States. Most often caused by the bacterium Borrelia burgdorferi, antibodies of the human adaptive immune system respond to proteins expressed by the bacterium to protect against the foreign pathogen. Current methods of testing for the presence of Lyme disease utilize immunofluorescence to detect the immune response to the outer surface proteins of Borrelia burgdorferi. This method lacks efficiency and accuracy because it can take several weeks to develop a detectable immune response and even then, the antibodies present may be in response to a different pathogen, leading to the possibility of false positives. The purpose of this research is to develop a magnetic bead-based enzyme-linked immunoassay to detect the IgM levels as the first adaptive response to Borrelia burgdorferi. We anticipate that the metallic beads will increase the sensitivity and efficiency of the ELISA because of the greater surface area, allowing for increased interaction between proteins. Through this research, an optimization of the capture protein concentration, detection antibody concentration and blocking conditions of the plate-based ELISA procedure was determined. Once a reasonable level of sensitivity was achieved, the ELISA was modified with a bead-based format with attachment of outer surface protein A to magnetic beads. We expected this technique to detect Lyme disease earlier and more accurately.

Development of a magnetic bead-based ELISA for detecting the early adaptive immune response to lyme disease

ACS Spring 2022

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