Tick-Borne Illness Diagnosis Enters New Era With Multiplex Testing

Co-infections from tick bites are often underdiagnosed due to several overlapping challenges. Many tick-borne illnesses – such as Lyme disease, Babesia, Anaplasma, Bartonella and Ehrlichia – share common symptoms like fatigue, fever, muscle aches and joint pain. These symptoms mimic those of viral infections or autoimmune conditions, making diagnosis difficult, especially in non-endemic regions.

Presentations of these diseases are complex and variable. Lyme disease may appear with a distinctive bull’s-eye rash, but many cases lack this hallmark. Other infections like Babesia or Bartonella often cause vague symptoms such as headaches or muscle pain, which are easily mistaken for other conditions. Co-infections further complicate diagnosis, as multiple pathogens can cause overlapping or intensified symptoms – joint pain from Lyme combined with Babesia-induced fatigue, for example – which may confuse clinicians and delay treatment.

Testing is another barrier. Standard diagnostic protocols focus primarily on Lyme disease using a two-tiered system — enzyme-linked immunosorbent assay (ELISA) and western blot — that fails to detect co-infections like Babesia or Bartonella. Testing for multiple pathogens requires specialized panels, which are not routine in most practices.

Additionally, many healthcare providers lack training or awareness regarding tick-borne co-infections. Even in Lyme-endemic areas, physicians may not consider other tick-borne diseases, and in non-endemic regions, these illnesses are often overlooked entirely. Finally, symptoms often appear gradually over weeks or months and patients may not recall a tick bite, making it less likely that providers will suspect a tick-borne illness during clinical evaluation.

Current testing methods can fall short in detecting multiple tick-borne pathogens simultaneously because they’re mostly designed to identify just one disease at a time – usually Lyme disease. The standard Centers for Disease Control and Prevention-recommended Lyme test, for example, uses a two-tiered method (ELISA followed by western blot) that’s not built to detect co-infections like Babesia, Bartonella or Anaplasma (amongst others).

Each pathogen requires a specific test. Many of these are not routinely ordered unless a doctor is highly specialized or the patient advocates for it. Patients with multiple infections often get partial diagnoses, or none at all, because the current system isn’t built to test everything at once – newer technologies aim to address this gap.

Traditional Lyme testing methods have significant limitations, especially in detecting early-stage disease or co-infections. The ELISA test, used for screening, can’t detect antibodies that take weeks to form. If ELISA is negative, testing usually stops – even if symptoms persist. Western blot results rely on the visual interpretation of protein bands, an outdated and inconsistent method that frequently yields false negatives.

Vibrant America’s chemiluminescent multiplex protein array offers a more sensitive, specific alternative, testing for multiple pathogens – including Borrelia, Babesia, Bartonella, Anaplasma and Ehrlichia – in one panel. Instead of visual analysis, it uses chemiluminescence immunoassay, enabling the detection of lower antibody concentrations, aiding early diagnosis and identifying subtle immune responses. By using highly specific peptides, the test also reduces cross-reactivity and improves accuracy.

To further enhance early-stage detection, Vibrant Wellness has developed an optimized immunoglobulin M (IgM) assay that removes excess immunoglobulin G (IgG) antibodies and rheumatoid factors from the serum. This process prevents elevated IgG from masking IgM, minimizing false negatives in the early phase of infection.

By enhancing sensitivity and specificity, Vibrant’s platform provides a more comprehensive diagnostic tool for complex, multi-pathogen, tick-borne illnesses. Antibody detection remains vital, as it reflects the immune system’s response — offering insight even after pathogens disappear.

In a 2020 peer-reviewed study published in Scientific Reports, researchers from Vibrant America demonstrated that their ultra-high-density peptide microarray platform offered marked improvements in both sensitivity and specificity compared to the traditional two-tier testing method for Lyme disease.

The study validated their single-tier assay across multiple patient cohorts and showed that the platform could accurately detect Borrelia-specific antibodies with greater reliability than conventional ELISA and western blot tests, especially in early and disseminated stages of infection. By using well-characterized peptides and chemiluminescent detection, the assay minimized cross-reactivity and reduced false negatives, establishing a more precise and scalable solution for tick-borne diagnostics.

This testing is available to clinicians. Providers can order the test for their patients and Vibrant offers a blood-spot version that requires a finger stick, which means it can be done at home without the need for a traditional blood draw. This can be ideal for patients who live in remote areas or have limited access to specialty care.

This at-home sample type is currently used for antibody-based detection only — we’re not able to run PCR on dried blood spots currently. As technology advances, I’m excited to explore ways to expand point-of-care and at-home options, with the goal of making comprehensive tick-borne disease testing more convenient, faster and widely available.

Multiplex testing has enormous potential far beyond tick-borne diseases. The ability to detect multiple pathogens simultaneously, with high sensitivity and specificity, could completely transform how we approach vector-borne and chronic infectious diseases on a broader scale.

For example, in areas where mosquitoes transmit dengue, chikungunya, Zika or malaria, symptoms often overlap – just like with tick-borne illnesses. A multiplex panel tailored for those regions could allow for rapid, accurate differential diagnosis, which is critical not just for individual patient care but also for outbreak control and public health surveillance.