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What Is Western Blot Test For Lyme

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What Abnormal Results Mean

Western Blot – Theory and method

A positive ELISA result is abnormal. This means antibodies were seen in your blood sample. But, this does not confirm a diagnosis of Lyme disease. A positive ELISA result must be followed up with a Western blot test. Only a positive Western blot test can confirm the diagnosis of Lyme disease.

For many people, the ELISA test remains positive, even after they have been treated for Lyme disease and no longer have symptoms.

A positive ELISA test may also occur with certain diseases not related to Lyme disease, such as rheumatoid arthritis.

The Igenex Lyme Immunoblot Solves These Problems

IGeneX has developed a serological test that increases specificity without sacrificing sensitivity that has changed how to test for Lyme disease. It uses specifically created recombinant proteins from multiple species and strains of Lyme borreliae and reduces inconsistencies in reading and interpreting the test bands.

More species detected The Lyme ImmunoBlot tests for more species of Lyme borreliae than the traditional ELISA and Western blot tests, reducing the risks of false negatives due to the inability to detect antibodies to a certain strain or species of Lb. The test includes all Borrelia-specific antigens relevant in North America and Europe, not just B. burgdorferi B31 or 297.

The result is a single test that replaces at least 8 Western blots.

More accurate testing The ImmunoBlot uses specific recombinant proteins that are sprayed in precise amounts onto specific locations on the membrane strip, allowing for greater control of the quantity and location of the antigens. This makes reading the bands much more accurate and consistent.

Earlier detection The ImmunoBlot can detect infections at multiple stages of illness, letting you catch infections earlier.

The IgM and IgG ImmunoBlots superior specificity and sensitivity make them the best Lyme disease test available.

Lyme Disease Testing: Useful When Performed Appropriately

One known problem is that we can produce antibodies for years or decades after a Lyme infection has been eradicated. Therefore, a seropositive test on its own is not necessarily indicative of active infection.

In addition, it can take a few weeks for detectable antibodies to build up in the body. Pseudoscience advocates frequently mislead about Lyme antibody testing by failing to differentiate testing performance in early infection from testing performance in late infection.

Below is a CDC-produced illustrative example of antibody production in Lyme disease, which triggers a positive test once a detection threshold is reached.

Two types of antibodies are shown: IgG and IgM. Typically, a detectable IgM antibody response is produced first, followed by a detectable IgG response.

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Indirect Testing Can Be More Limited Than Direct Testing

Both tests used in the CDC recommended two-step process are indirect methods of diagnosismeaning they do not detect the actual Lyme disease bacteria itself but, instead, measure the bodys immune system response to the presence of disease-causing bacteria. However, a number of factors can prevent the body from producing antibodies, including the following:

  • Timing: If conducted in the early stages of Lyme disease, a patients body may not have developed a sufficient enough number of antibodies to detect. This issue can be compounded by the lack of sensitivity of the ELISA test.
  • Immunity Suppression: The saliva of infected ticks contains specific immune-suppressing components that can delay or prevent the activation of a persons immune response. These components are designed to prevent the body from effectively fighting off the Lyme disease-causing bacteria, so it has a chance to take hold.
  • Antibiotics: If patients are taking antibiotics at the time of the Lyme disease test, they may not produce enough antibodies to be detected by the test.
  • Modified Form of Borrelia: In some patients, the Borrelia bacteria will transform into a cyst, which will prevent the bodys immune system from producing antibodies.
  • Weakened Immune System: False negatives can also result in patients in whom the immune system is weakened or compromised due to coinfection with another illness.
  • Seronegative patients: These patients do not produce antibodies.

References Choosing A Test

When Life Gives You Lyme: ELISA &  Western Blot Tests for Dummies
  • Wormser GP, Liveris D, Hanincova K et al. Effect of Borrelia burgdorferi genotype on the sensitivity of C6 and 2-tier testing in North American patients with culture-confirmed Lyme disease. Clin Infect Dis, 47, 910-914 .
  • Wormser GP, Schriefer M, Aguero-Rosenfeld ME et al. Single-tier testing with the C6 peptide ELISA kit compared with two-tier testing for Lyme disease. Diagn Microbiol Infect Dis, .
  • Read Also: Signs Of Lyme Disease In People

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    Lyme Disease Testing: Why It Continues To Fail To Properly Identify Lyme Disease

    Darin Ingels, ND, FAEM

    Dr. Ingels spoke at a past Restorative Medicine Conference and is on the Scientific Advisory Board of the Restorative Medicine Herbal Certification program.

    When I was working as a microbiologist in the early 1990s, Lyme testing was still pretty new and we didnt know much about Lyme disease and all of the nuances of testing that would soon follow. Now that it is 25 years later, we have learned a great deal about Lyme disease and the pitfalls of Lyme testing have been revealed. Here is a brief excerpt from my upcoming book on Lyme disease, The Lyme Solution: A 5-part Plan to Fight the Inflammatory Auto-Immune Response and Beat Lyme Disease.

    The current recommendation by the CDC for someone who is suspicious of having Lyme disease is to run a two-step process. The first step is to run a Lyme antibody blood test, which measure two types of antibodies against Lyme disease . If this test is positive, then a second test called a Lyme Western Blot should be run to confirm the results of the first test. The Lyme Western Blot looks at a series of specific antibodies against the Lyme organism for both IgG and IgM. If someone has at least 5 out of 10 antibodies for IgG or 2 out of 3 antibodies for IgM, then the Lyme Western Blot is considered positive. The CDC does not recommend doing a Lyme Western Blot IgM test on anyone whose illness occurred within the past month.

    Stony Brook University Medical Center

    Administered in conjunction with the Clinical Immunology Laboratory in the Department of Pathology at the Renaissance School of Medicine at Stony Brook University, the IgM and IgG Western Blot Lyme disease test is often covered by insurance. This is considered a more sensitive test if the ordering healthcare provider checks the box to include CDC non-specific bands on the Western Blot report.

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    And Heres Where It Gets Interesting

    Around the time that the CDC was making its decision about the classifications, GSK was trying to develop a Lyme vaccine. The point of a vaccine is to develop antibodies to a certain part of the infection, and the GSK vaccine was, in fact, creating antibody responses in patients that would create a positive for bands 31 and 34 on the Western blot. In other words, anyone who received the Lyme vaccine would have a positive Western blot if they included bands 31 and 34 in the official criteria.

    So those bands were scrapped in order to avoid what they thought might be a bunch of false positives in recipients of the vaccine. The problem is, so was the vaccine! Very few people ever got it because of the side effects. But the CDC never revised their criteria, despite plenty of science to the contrary.

    How To Read Western Blot Test Results

    Lyme Disease: POSITIVE IGM Western Blot Test (2013)

    The Western Blot Test does not actually test for the presence of Lyme disease. It tests indirectly by looking for antibodies made by your immune system agains Lyme disease bacteria, spirochetes, which causes Lyme disease.

    Western Blot Bands description:

    Cross-reactive means it could indicate other spirochetes.

    Specific means it only happnes when Lyme disease is present.

    Borellia or Bb is Lyme disease.


    22 specific for Bb, probably really the 23/25 band23-25 outer surface protein C , specific for Bb 28 outer surface protein D Oms28 specific for Bb 30 OspA substrate binding protein common in European and one California strain31 outer surface protein A , specific for Bb34 outer surface protein B specific for Bb35 specific for Bb38 cross-reactive for Bb39 is a major protein of Bb flagellin specific for Bb41 flagellin protein of all spirochetes this is usually the first to appear after a Bb infection45 cross-reactive for all Borellia 50 cross-reactive for all Borrellia55 cross-reactive for all Borrellia57 cross-reactive for all Borrellia58 unknown but may be a heat-shock Bb protein60 cross reactive for all Borrellia66 cross-reactive for all Borrelia, common in all bacteria 83 specific antigen for the Lyme bacterium, probably a cytoplasmic membrane93 unknown, probably the same protein in band 83, just migrates differently in some patients

    The bands highlighted in RED above are very specific for Lyme disease.

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    Should You Get Tested For Lyme Disease

    The current laboratory diagnostic tests for Lyme disease are blood tests to detect the antibodies created by your body to fight and kill the bacteria that causes Lyme. This bacteria is called Borrelia burgdorferi, or Bb.

    However, with early-stage Lyme your antibodies may not have had time to develop in your body. Therefore a blood test at this time will often appear negative. It can take your body up to two months to develop enough antibodies to be detected by these tests. In other words you could have Lyme but the test results will still come up negative.

    What does this mean for you? By all means, if your doctor recommends a test, get the test. But dont rely on it completely. Trust your body and trust your homework. Monitor your symptoms. Ask questions.

    Lyme Disease Testing: How It Works

    Two common lab tests for Lyme disease are the enzyme immunoassay test and the Western blot test. These tests measure the antibodies that form when someone is infected with Borrelia burgdorferi .

    Enzyme immunoassay test

    The EIA test is whatâs typically used to screen for Lyme disease by detecting antibodies for B. burgdorferi.

    According to the Centers for Disease Control and Prevention, if the EIA test shows negative for the antibodies, no further testing is needed. However, if this screening test comes back with results that are positive or unclear, a second round of testingâusing the Western blot methodâis recommended. This second test is referred to as a confirmatory test.

    Western blot test

    When the EIA test is positive, the Western blot test is usually done next. This two-step approach to Lyme disease testing helps reduce the number of false positives. The Western blot test detects antibodies for several proteins belonging to Lyme bacteria.

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    Experts Agree Around The World

    A strong scientific consensus is evident about Lyme disease diagnosis and testing.

    A 2018 French review of 16 Lyme diagnostic guidelines from 7 countries revealed a global consensus regarding diagnosis at each stage of the infection. The only outlier was the pseudoscience group German Borreliosis Society , a German counterpart to the pseudoscience group ILADS.

    Predatory Labs: How They Trick People

    Lyme diagnosis and lab results  Lyme Tribune

    Experts strongly warn against unvalidated testing . Quackwatch and Science-Based Medicine both provide accessible explanations about the differences between validated, science-based tests and unvalidated tests.

    In North America, only CDC-recommended and FDA-cleared or approved testing should be used. Health agencies in Canada make the same recommendations as CDC.

    Sadly, a number of predatory labs exploit permissive laws surrounding lab developed tests to peddle testing that should not be relied upon.

    Some predatory labs boast that they are CLIA-certified, but such lab certification does not mean that a test is useful. Even the notoriously fraudulent lab Theranos was CLIA certified.

    CDC scientists explain:

    • CLIA certification of a laboratory indicates that the laboratory meets a set of basic quality standards.
    • It is important to note, however, that the CLIA program does not address the clinical validity of a specific test .
    • FDA clearance/approval of a test, on the other hand, provides assurance that the test itself has adequate analytical and clinical validation and is safe and effective.

    Besides misrepresenting CLIA certification, predatory labs use other methods to trick patients, including:

    • Charging big bucks,
    • Testing for many biomarkers to maximize the chance of a non-negative result,
    • Creating proprietary diagnostic criteria, and
    • Returning indeterminate results, which will often be interpreted as positive.

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    Lyme Disease Tests: Science Vs Misconceptions

    Lyme diagnosis and testing can be very confusing. This confusion has contributed to false positive diagnoses and unnecessary treatment.

    As in other infections like HIV, testing for Lyme disease involves looking for antibodies produced by the bodys immune system in response to infection. This is called serologic testing because the antibodies are found in blood serum.

    Testing positive for antibodies is called seropositive and testing negative for antibodies is called seronegative.

    More About Western Blot Testing In North America

    The IgG Western Blot test is designed to detect antibodies specific to Borrelia burgdorferi, the bacteria that cause Lyme disease. For Lyme disease in North America, a positive IgG Western Blot test requires at least 5 of 10 measured bands to be positive .

    The scored IgG bands are 18 kDa, 24 kDa , 28 kDa, 30 kDa, 39 kDa , 41 kDa flagellin , 45 kDa, 58 kDa , 66 kDa, and 93 kDa.

    The Lyme IgM Western Blot test measures 3 different types of antibodies. The North American IgM Western Blot is considered positive only if 2 of 3 IgM bands are positive .

    The scored IgM bands are 24 kDa , 39 kDa , and 41 kDa .

    As discussed above, the IgM Western Blot should only be used in the first 30 days of illness.

    According to CDC, Depending upon the assay, OspC could be indicated by a band of 21, 22, 23, 24 or 25 kDA.

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    Lyme Disease Immunoblot Test

    The Lyme Disease Immunoblot test is typically used as a confirmatory test for people who have had positive results from previous Lyme Disease testing. The immunoblot test is also known as a Lyme Disease Line Blot test. This test looks for the presence of both IgG and IgM antigen bands to confirm both recent and previous exposure. The detection of multiple bands is required for a positive result. The CDC does not recommend the Western Blot test as a front-line screening as some conditions other than Lyme Disease may cause a false positive.

    Lyme Disease is caused by infection with the bacteria Borrelia burgdorferi. It is most commonly spread by the Deer Tick or Black Legged Tick and is the most common tick-borne illness in the United States and Europe. Lyme disease can be difficult to diagnose because the symptoms are often similar to other afflictions. While some people will display a distinctive “bulls-eye” rash around the site of the bite, others may experience common flu-like symptoms such as fever, chills, fatigue, and headache. The ticks which spread the disease are typically very small and many people do not even realize they have been bitten. If left untreated, Lyme Disease can cause joint pain, numbness in the arms and legs, facial paralysis, Meningitis, heart problems, or memory problems.

    Turnaround time for the Lyme Disease Immunoblot test is typically 2-5 business days.

    Detection Period:

    How Is Lyme Disease Diagnosed

    The Western Blot Test And Lyme

    Though several types of tests do exist for the diagnosis of Lyme disease, the best tests for a Lyme disease diagnosis are blood tests, also known as serological tests. These tests are indirect, meaning they dont detect the infecting bacteria or its antigens but rather the antibodies an infected persons body produces in response to these antigens.

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    What Is The Western Blot Test Used For

    The Western blot test is used to confirm or contest a diagnosis of HIV or Lyme disease after an ELISA antibody test comes back positive or negative. Since the ELISA test sometimes produces false positives, a second test is needed to further the diagnosis.

    If you have Lyme disease, the Western blot test may detect antibodies to B. burgdorferi, the bacteria that causes this condition.

    If you have HIV, the Western blot test may detect proteins from the envelope or core of the virus and enzymes generated by HIV infection.

    The Western blot test is a simple blood test. It doesnt require fasting, or preparation in any way.

    Like all blood tests, you may need to fill out a consent form first.

    You may have blood drawn at your doctors office thats sent to a lab, or you may go directly to the lab for testing once the test has been ordered. Based upon your doctors protocols and the lab used, you should have your test results in 4 or 5 business days.

    Just like all blood tests, there are no serious side effects to expect. You may have a slight amount of bruising at the blood draw site. If you get nervous or are uncomfortable doing blood draws, it may help to bring someone with you.

    It can also be stressful or worrisome to wait for test results. Keep in mind that testing is a tool that helps your medical team care for you and provide treatments that can manage your condition.

    When Is The Lyme Disease Blood Test Ordered

    When someone has the signs and symptoms of Lyme disease or they live in a region that has deer ticks or black-legged ticks, then this blood test will be ordered. It will also be ordered when these symptoms occur without improvement over the course of 7-14 days by most medical providers.

    • A bulls-eye rash that grows from the bite site.
    • Fevers, chills, and a persistent headache that does not go away.
    • Unusual and persistent fatigue that does not immediately improve.

    The IgM and IgG tests are generally ordered first when Lyme disease is suspected. This is because people who have never been exposed to the bacteria that causes the disease will not have any antibodies present. If these tests are positive and followed up by a positive Western Blot test, then the chances are very good that Lyme disease is present. This is especially true if antibody levels continue to rise over time.

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