Developing a Novel Species-Independent Biosensor for Lyme Disease via the Antibody-Catalyzed Water Oxidation Pathway

Abstract

Lyme disease, also known as Lyme Borreliosis, is caused by bacteria of the Borrelia type, and it is the most common vector-borne infectious disease spread by ticks in the densely populated Northern Hemisphere including Asia, Europe, and North America. Lyme disease can lead to significant pains and sufferings, making it one of the most insufferable diseases. If left untreated, infected patients can develop symptoms as serious as paralysis, chronic shooting pain, heart problems among others. Early detection is essential in proper Lyme disease treatment. The current technique used to detect Lyme disease is a two-step approach including an ELISA (enzyme-linked immunosorbent assay) test and a confirmatory Western blot test. However, ELISA is not an ideal test because it relies on the use of secondary antibodies to detect primary antibodies. Due to non-specific binding of secondary antibodies, ELISA suffers from a high false-positive rate, making a positive result unreliable and must be confirmed by an additional Western blot test. In addition, since secondary antibodies are specific to species, a different Lyme test protocol is required for each different species. Lastly, because secondary antibodies are not cheap and not always readily available, an ELISA-based Lyme test is unnecessarily expensive and slow. There is, however, a novel technique that overcomes the disadvantages of the ELISA-based Lyme test. By using the antibody-catalyzed water oxidation pathway (ACWOP), a novel biosensor can be designed to directly detect the presence of primary antibodies to Borrelia without the use of enzyme-tagged secondary antibodies. In ACWOP, the intrinsic catalytic power of all antibodies regardless of species is utilized to oxidize water into hydrogen peroxide for detection. Addition of readout reagents in the presence of hydrogen peroxide can then generate either colorimetric or fluorometric signals that can be easily detected. Since ACWOP eliminates the use of species-specific secondary antibodies, the novel Lyme test based on ACWOP will have higher specificity, more convenience and more affordability, and most importantly, unify human and veterinary serological testing.