LYME DISEASE (UPCOMING)

Guanine will offer the first mobile Lyme test with high accuracy, AI assessment, AI diagnosis and telehealth option

Individuals can validate suspicious rashes and symptoms using AI tools that reduce misdiagnosis by expanding the criteria for ruling-in suspicious infections for testing and for differential diagnosis after test results

Individuals can get a Guanine test with results in minutes at a test center or with a smart phone adapter for self testing

AI  using symptoms and test results encourages individuals diagnosed positive for Lyme disease to get seen by a healthcare professional for confirmation and treatment.

Why is Lyme disease so difficult to detect and diagnose?

Lyme disease diagnosis is complex and lengthy. Lyme disease symptoms can appear quickly or gradually over time and are incredibly varied. The first physical signs of Lyme disease are often flu-like symptoms which could be confused with the flu or the common cold. Erythema migrans (EM) skin rash is the most common manifestation of Lyme disease with up to 80% of infected persons developing a variation of this skin lesion in the first weeks of infection.

• EM can have 5 forms but the bull’s eye EM manifests in only about 20% of patients with EM, with the majority of EM lacking the central clearing or ring-within-a-ring bull’s eye pattern.
• One study showed that 72% of physicians surveyed were unable to correctly identify the EM when shown both EM and other rashes common in an ambulatory population.
• Another study showed that 72% of physicians misdiagnosed skin lesions as EM before EM forms (i.e. < 5 days from the tick bite) or after the EM has faded (i.e. > 30 days post tick bite).

Borrelia burgdorferi bacteria which cause Lyme disease have a spiral shape and penetrate human tissue like a corkscrew to enter the bloodstream, use up to 10 flagella to speed through the bloodstream, and survive outside of the bloodstream because they do not require iron. Unlike sepsis-causing bacteria, Borrelia does not stay in the blood for a long time.

• Direct testing for Borrelia have variable sensitivity (i.e. true positive rates) including culture (27% - 94%) and PCR (34% - 62%) since Borrelia may not be present if the sample is collected at the wrong time.

Patients infected by Borrelia produce antibodies which are used to detect Lyme disease becuase of the variability and high cost of directly detecting Borrelia. In 1994, a 2-tiered antibody testing strategy was adopted by the CDC in which a first-tier ELISA test was used to detect possible cases, and a second-tier test Western blot was added as a more specific confirmatory step to rule out false positives.

• However the two-tier test incurred many shortcomings including a time lag for the host to produce detectable antibody quantities. This resulted in a true positive rate of only 46% in acute Lyme disease detection.
• The true positive rate was lower in patients with weakened immune systems who were more likely to incur chronic Lyme disease.
• The two-tier test is also unable to distinguish an active infection or re-infection from previous exposure.

Other tickborne pathogens are increasing transmitted with Borrelia including bacteria (Bartonella, Rickettsia, Mycoplasma), protozoa (Babesia), and viruses (Powassan) which can require different treatment to resolve and different tests to detect.

• Co-infections are not tested very often.

Guanine will improve acute Lyme disease test sensitivity from 46% to 85-95%

The standard two-tier testing (STTT) process employs an ELISA antibody screening test followed by a Western Blot confirmation test for IgM proteins. STTT test performance for acute Lyme disease detects only 46% of infected patients. As a consequence, the majority of Lyme disease patients are not detected by testing in the acute stage and incur late stage Lyme disease which is more difficult and expensive to treat.

Lyme disease patients produce a serological biomarker profile comprising a Borrelia burgdorferi (Bb) peak about 2-9 days post tickbite, an IgM peak about 16-28 days post tickbite, and a IgG plateau that exceeds IgM days after IgM declines from its peak. Direct detection of Bb is an effective indicator of Lyme disease, however, Bb does not remain in the blood for a long duration making post tickbite testing time highly variable.

Lyme disease is confirmed by the presence of 2 of 3 IgM proteins as visible bands. One of these proteins p41 is associated with the Borrelia flagella which is common to other microbes such as syphilis bacteria as is a source of false negative tests for Lyme disease. WB has a limit of detection (LOD) of about 10^8 analytes/mL, compared with about 10^6 for ELISA.

As a consequence, patients with low antibody production rates (i.e. immuno-suppressed, HIV, elderly, patients taking certain medications), are unable to attain IgM production of 10^8 analytes/mL and produce a false negative test result. IgG is a more specific biomarker than IgM however, it can remain in the blood for years after an infection, making positive IgG antibodies unable to distinguish a new infection from a previous infection.

Guanine's Lyme disease test

Guanine's Lyme disease test will detect IgM and IgG antibodies, and Borrelia burgdorferi bacteria from the same test sample on three separate electrodes.

ELECTRODE 1 - IgM
Three IgM antibody proteins (p21, p39, p41) will be individually quantified at 4 logs lower than Western Blot to reduce false negatives from infected people with low antibody production rates. Nonspecific materials will be removed with magnetic separation to reduce false positives.

ELECTRODE 2 - IgG
IgG will be quantified to determine if the patient was previously infected based on the IgM/IgG ratio.

ELECTRODE 3 - Borrelia Bacteria
Three different nucleic acid targets from the Borrelia burgdorferi bacteria will be detected using RNA, OspC gene and VlsE gene to reduce false negatives before measurable antibody production.  Borrelia is not detected in the current standard two-tier testing and reduces sensitivity.

The Lyme disease test will be expanded to detect other tick co-infections from the same sample.

In addition to testing, individuals can validate suspicious rashes and symptoms using AI tools that reduce misdiagnosis by expanding the criteria for ruling-in suspicious infections for testing and for differential diagnosis after test results.