Best Test for Diagnosing Lyme Disease

When it comes to diagnosing Lyme disease, the best test for Lyme disease uses a combination of blood work from standard lab tests as well as physician interpretation and experience. In certain circumstances we turn to labs that perform specialty testing for Lyme and other co-infections.

In this post we will discuss the best test for Lyme disease; including specialty tests we choose, when we use these tests, and why we believe these tests are necessary.

As a rheumatology clinic, we see quite a few patients suffering from chronic infections, including Lyme disease. Since 2009, we have diagnosed and treated several thousand patients with chronic Lyme disease.

Diagnosing Lyme disease: Laboratory Testing and Physician Interpretation
Because Lyme disease is a chronic infection similar to many chronic infections, the nature of Lyme disease is characterized by periods of exacerbations and remission. As a physician, when I suspect Lyme disease as the cause of illness, there are three key questions I am looking to find answers to with blood work.

These questions are:

  1. Does the patient have Lyme disease?
  2. Is the patient dealing with acute Lyme disease or is the disease in a dormant state?
  3. What other co-infection(s) can be present?

Diagnosing Acute Lyme disease
The laboratory diagnosis of acute Lyme disease can occur from several days up to 3-4 weeks from the tick bite episode. It is based on detection of microorganisms, their antigens, or nucleic acids in the tissues (for example, the skin) and biological fluids (blood, cerebrospinal fluid, synovial fluid, saliva, and urine), as well as detection of Borrelia-specific IgM antibodies in the blood.

An antibody, or immunoglobulin, is a protein that your immune system produces to neutralize foreign materials such as bacteria and viruses. Therefore, if high levels of antibodies are detected in your blood, it is a good indication that an infection is present.

The body makes different immunoglobulins to combat different viruses. There are five subclasses of antibodies, IgA, IgG, IgM, IgE, and IgD.

In general, the presence of either IgG or IgM antibodies to the Borrelia burgdorferi (Borrelia burgdorferi is the bacteria that causes Lyme disease) protein is indicative for Lyme disease. IgM is the first immunoglobulin produced following exposure to a virus; therefore, in acute cases IgM will be present. IgG antibodies remember what viruses you have been exposed to and if they come back, they are produced again.

So testing for IgG lets you know if you have been infected before. IgG is helpful in diagnosing chronic Lyme disease—meaning someone has been infected for a while.

What are the Best Tests for Lyme Disease?

Diagnosing Chronic Lyme disease: The Western Blot Test
Typically, the standard for determining the presence of chronic Lyme disease is confirmed by the Lyme Western Blot blood test. This test can detect two different classes of antibodies: IgM and IgG.

During the procedure proteins extracted from Borrelia burgdorferi microbial cells and a patient’s blood are placed onto gel strips where they can form protein bands. If a patient’s blood contains antibodies to Borrelia burgdorferi, they react with the bands in the gel forming so called “positive bands.”

The CDC requires 5 out of 10 bands for a positive test result. However, because some bands on the Western blot are more significant than others your doctor may decide you have Lyme disease even if your Western blot does not have the number of bands or specific bands recommended by the CDC.

Some Lyme specialists and scientists believe that there are five very specific bands on the Western blot test that are highly indicative of Lyme disease, band numbers 23, 31, 34, 39 and 93. If any of these bands are positive and the patient is experiencing symptoms of Lyme disease, they may feel treatment is warranted.

In addition, the test systems on the market are not standardized. Therefore, findings from different laboratories can be compared to only a limited degree. Tests focused on detection of antibodies to other tick-borne microorganisms or their nucleic acids (other borrelia species, bartonella, and babesia species, etc.) sometimes require overlapping testing through different laboratories to confirm the diagnosis.

If the physician suspects Lyme, but the patient has an inconclusive Lyme Western Blot test result, another approach can be used. In our clinic we call it band shift.

Let’s presume that a patient with a clinical picture suspicious for Lyme disease has only one or few bands in the Lyme Western Blot, which is/are not the most unique or specific for Borrelia burgdorferi.

In this situation, we ask the patient to take an antibiotic or antimicrobial herb for 1-2 months, and then repeat the Western Blot test. If there is a change in the band profile, to the highly specific bands, then this is an indication the disease is present.

Why does this happen? Taking an antibiotic or antimicrobial kills the Borrelia pathogen and releases toxins into your bloodstream. In response to the release of toxins, your antibodies increase, resulting in the band shift.

The Western Blot test measures for levels of antibodies to the Borrelia, not the Borrelia itself.

At IFSMED we also perform Lyme Western blot on synovial fluid (fluid aspirated from swollen joints)

Our joints and synovial spaces represent biological traps for various microbioligical antigens which stimulate a local intrasynovial immune response. Based on that principle, synovial fluid analysis should produce more positive results than the corresponding serum analysis.

Based on our statistics, patients with Lyme disease have more positive bands upon analysis of synovial fluid as compared to the paired serum samples.

This image is from the same patient demonstrating more positive bands with synovial fluid versus serum. Synovial fluid test results on the left, blood test results on the right. 

serum versus synovial lyme results


A positive Lyme Western Blot means that the patient has acquired a Borrelia infection at some point in time. With a single serological test it is not possible to decide whether this infection is active or dormant. It is important to understand that a negative serological finding does not rule out Lyme disease.

In our clinic, we use many different factors to diagnose Lyme disease including the following laboratory markers to best test for Lyme disease activity.

None of these markers are unique for Lyme disease. However, taken together with clinical symptoms and Lyme Western Blot results, they can be used as surrogate markers of disease activity. Furthermore, these markers can be used to monitor the outcome of antimicrobial therapy in patients with Lyme disease.

C4a: This marker serves as a mediator of the local inflammatory process and is commonly highly elevated among people with Lyme disease. It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes. Patients with active Lyme disease are characterized by the increased blood levels of C4a.

CD57: is a marker present on natural killer (NK) cells. NK cells are activated as a response to an infection and work to control infections. Patients with chronic Lyme disease are characterized by a low count of CD57 + NK cells compared to those with acute Lyme disease.

Immune complexes: An immune complex, sometimes called an antigen-antibody complex, is a molecule formed from the integral binding of an antibody to a soluble antigen. An increase in the concentration of immune complexes is found in patients with various chronic infections, including Lyme disease, as well as in patients with certain autoimmune disease, for example, systemic lupus erythematosis.

IgM antibodies to phospholipids (APL antibodies): The presence of low affinity IgM APL antibodies is seen in various chronic infections, including chronic Lyme disease. Affinity is the binding strength of the antibody to an antigen. It determines the rate at which an infection is eliminated. The higher the affinity, the faster an infection will be eliminated.

Anti-histone and anti –chromatin antibodies: are autoantibodies frequently found in patients with autoimmune and chronic infectious diseases, including Lyme disease. Most likely, the level of these antibodies parallels the concentration of immune complexes.

Specialty labs for Lyme disease tests—are they necessary?

As an alternative to standard Western Blot tests, in certain instances we also use specialty lab tests where their individual focus is on developing and administering the most precise and effective diagnostic tests for Lyme disease and other tick-borne diseases.

When should someone consider using a specialty lab as opposed to standard blood work?

Typically, we use reference labs to confirm various co-infections or to test antibodies against tick-borne microorganisms in biological fluids different from the blood (for example, synovial fluid – fluid from the swollen joints)

The following labs are tests we use on a regular basis. We will cover why we choose the tests we do, show you an example of each test, and when we believe additional testing is necessary.


IgeneX uses an ImmunoBlot test that detects antibodies to more B. burgdorferi species than the current Western blot does. By combining the results from more significant protein bands, IGeneX testing provides higher sensitivity for detecting Lyme exposure.

In fact, the ImmunoBlot test for Lyme disease has a sensitivity greater than 93%, whereas the ELISA and Western Blot two-tier testing protocol recommended by the CDC has a sensitivity of only 57.6%

Igenex claims to test more relevant strains of tick-borne pathogens than any other lab, allowing for one test to show results from many different species.

igenex igg igm criteria

best test for lyme disease igenex results
Igenex test result


Galaxy lab we use for Bartonella testing.

Their process uses PCR testing. Polymerase chain reaction, or PCR, is a method of detecting target bacterial DNA in a sample.

Research shows that the Bartonella bacteria that make people sick can cause a cyclical bacteremia. That means that sometimes the bacteria is in the blood and sometimes it isn’t. This allows the bacteria to evade the host immune response and makes it very hard to test for.

In order to overcome this challenge, Galaxy has developed a method of adding an enrichment culture to the sample allowing the bacteria to grow to detectable levels. Performing PCR on a sample after allowing bacteria to grow maximizes the chance of detecting present DNA. Using PCR after culturing reduces the chance of false negatives.

bartonella epcr explanation

DNA Connexions

Unlike the other tests, DNA Connexions uses a urine sample that you collect at home.

Prior to collecting, physical provocation is highly recommended.

This can include:

  • 30 minutes of vigorous exercise
  • 1 hour lymphatic or deep tissue massage
  • 30 minutes of infrared sauna

Why urine?
The urinary tract is known to be a place where spirochetes collect.

Why exercise first?
Many clinicians believe that exercise, infrared sauna, and deep tissue massage are good provocators of the microorganisms into the blood and then ultimately into the urine prior to urine collection.

This is because B. burgdorferi disseminates throughout the body and can hide in different places, such as the lymph nodes and joints.

Physical movement aids in releasing the organisms and getting them moving through the system, including the urinary tract.

dna connexions test result

Important considerations when searching for the best test for Lyme disease:

Different labs use different approaches, analytes and methods to diagnose a particular infection. Therefore, it is not uncommon to get different results from different labs targeting the same infection.

Remember, lab tests are an adjunct tool. The actual diagnosis is made by an experienced physician who relies not only on labs but on clinical symptoms, instrumental analysis and a patient’s history. It is not uncommon to make a diagnosis even in the absence of confirmatory lab test results.