Why Is It Called Western Blot
W. … Burnette definitely gave the technique the name “Western blotting” as a nod to Southern blotting and because their lab was on the west coast. He developed his technique independently, including the electrophoretic transfer step, but became aware of Stark’s and Towbin’s publications before he submitted his in 1979.
Testing For Lyme Disease Years Later
When a person doesnt know that theyve had Lyme for years and begins presenting with symptoms of a chronic Lyme disease infection, they will likely be tested to look for antibodies. This is because antibodies continue to build up within the system while the infection runs rampant.
Image by on How accurate are the results when testing for Lyme disease years later?
What Is The Most Reliable Lyme Disease Test
There is no one answer when it comes to determining which Lyme disease test is the most reliable. This is because each test looks for a different thing at a different stage of the infection. When investigating a long-term infection, tests that look for the bodys response to the infection are the most reliable, whereas for an active infection, a test that identifies the bacteria itself will be the most effective. Some tests only show up on an active infection and fail to indicate chronic Lyme, so the most reliable test option depends on which stage a persons infection is currently at.
Featured image by Emin Baycan on Unsplash
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How Is It Diagnosed
If youâve been outside in an area where ticks are known to live, you should tell your doctor. Theyâll also want to know about the symptoms youâre having. These details are crucial to making a diagnosis of Lyme disease.
Early symptoms that usually occur within the first month after a tick bite can include:
- Rash at the site of the tick bite that may look like a âbullâs-eyeâ
- Racing heart
- Problems with your short-term memory
Symptoms that come and go are common with Lyme disease. They will also depend on the stage of the disease.
Lyme Western Blot Igg
The IgG Western Blot is an immunoassay and qualitative test in which antibodies are visualized. The IgG antibody is typically present a few months following initial infection.
Clinical Significance
The Lyme Western Blot IgG is a sensitive indicator of an exposure to B. burgdorferi. IgG antibody is typically present a few months following the initial infection. Lyme Western Blot IgG must be performed on any sample with positive or equivocal result for Lyme IgG/IgM antibody serology or Lyme IgG/IgA/IgM IFA.
Limitations
Presence of an indeterminate number of double starred bands in a negative report may indicate clinical significance. Therefore, we recommend testing with another method and/or retesting in 4-6 weeks.
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Polymerase Chain Reaction May Help In Some Situations
Molecular assays are not part of the standard evaluation and should be used only in conjunction with serologic testing. These tests have high specificity but lack consistent sensitivity.
That said, PCR testing may be useful:
-
In early infection, before antibody responses develop
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In reinfection, when serologic tests are not reliable because the antibodies persist for many years after an infection in many patients
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In endemic areas where serologic testing has high false-positive rates due to high baseline population seropositivity for anti-Borrelia antibodies caused by subclinical infection.
PCR assays that target plasmid-borne genes encoding outer surface proteins A and C and VisE are more sensitive than those that detect chromosomal 16s ribosomal ribonucleic acid genes, as plasmid-rich blebs are shed in larger concentrations than chromosomal DNA during active infection. However, these plasmid-contained genes persist in body tissues and fluids even after the infection is cleared, and their detection may not necessarily correlate with ongoing disease. Detection of chromosomal 16s rRNA genes is a better predictor of true organism viability.
The disadvantage of PCR is that a positive result does not always mean active infection, as the DNA of the dead microbe persists for several months even after successful treatment.
How Lyme Antibody Testing Works
Antibody testing for Lyme disease requires two different tests to establish a positive result. If either the first tier test or the second tier test is negative, the test result is negative overall.
But in the event of a negative result, Dr. Adriana Marques of the NIH states:
For patients with signs or symptoms consistent with Lyme disease for less than or equal to 30 days, the provider may treat the patient and follow up with testing of convalescent-phase serum.
The first tier of the two-tiered testing system is an Enzyme Immunoassay .
The second tier of the well-established Standard Two-Tiered Testing involves a Western Blot test, which can be complicated to understand. The Western Blot is also called an immunoblot or a line blot.
The Western Blot test typically reports two types of antibodies that may be indicative of a Lyme infection: IgM and IgG.
According to the CDC, Positive IgM results should be disregarded if the patient has been ill for more than 30 days.
According to a consensus of experts, including representatives of the American Academy of Pediatrics, American Academy of Neurology, American College of Rheumatology, and Infectious Diseases Society of America:
Immunoglobulin G seronegativity in an untreated patient with months to years of symptoms essentially rules out the diagnosis of Lyme disease, barring laboratory error or a rare humoral immunodeficiency state.
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Lyme Disease Antibody Test Procedure
The Lyme disease antibody test requires no advance preparation. A lab technician will swab the inside of your elbow with an antiseptic before drawing your blood. Your blood will be drawn from a vein in your arm using a small needle.
The blood draw should not be painful, though you might feel a slight prick when the needle is inserted into your vein.
The blood sample will be collected in a vial. The puncture site will be bandaged, if needed, after the needle is removed. After the blood draw, you are free to go home.
There are very few risks associated with the Lyme disease antibody test. Excessive bleeding is possible, but there may be an increased risk if you take blood thinning medications or certain anti-inflammatory drugs like:
- heparin
How Accurate Are Lyme Disease Tests
Medically reviewed by Neka Miller, PhD on August 11, 2020. To give you technically accurate, evidence-based information, content published on the Everlywell blog is reviewed by credentialed professionals with expertise in medical and bioscience fields.
Interested in getting tested for Lyme disease and want to learn more about the testing process? Here, weâll cover several aspects of Lyme disease testing, including accuracy in the context of false negatives, different test types, and moreâso continue reading.
Check for Lyme disease from the convenience of home with the Everlywell at-home Lyme Disease Testâwhich includes both screening and confirmatory testing .
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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.
Lyme Disease Blood Test Results Fully Explained
The Lyme disease blood test is used to discover if someone who has the symptoms of a Borrelia burgdorferi infection actually has the bacteria in their bloodstream. Recent infections are much easier to detect and an IgM and IgG blood test will often be ordered as complimentary information gathering tools. This blood test does not always detect the presence of the disease, so patients that have persistent symptoms after having the test may be re-tested in a few weeks.
If any of these tests come back as positive, then other samples will be used to track the stage of the disease to determine if it has reached the chronic infection stage. At this point, a medical provider will order a Western blot test to confirm the presence of Lyme disease.
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Lyme Disease Test Two
Two-tiered Lyme disease testing uses two tests. The first is a screening test that should detect anyone who might have the disease. Tests that do this well have are regarded as having high sensitivity. This test is followed by a second test that is intended to make sure that only people with the disease are diagnosed. Tests that do this well have high specificity.
HIV/AIDS is diagnosed with tests that are both highly sensitive and highly specific. They are accurate more than 99% of the time. In Lyme disease, the second test is highly specific. So there are few false positives. Unfortunately, the screening test is highly insensitive and fails to accurately identify patients who have Lyme disease. The two-tiered test system misses roughly 54% of patients.
Because of this, LDo recommends the patients and physicians skip the ELISA and go straight to the Western blot.
What To Think About
- It may be hard to tell if you have Lyme disease. False-positive and false-negative Lyme disease test results are common. Many people do not make antibodies to Lyme disease bacteria for up to 8 weeks after being infected.
- Doctors often do not rely on test results alone when recommending treatment for a person who may have Lyme disease. Treatment is often based on a person’s symptoms, the time of year, having a tick bite, and other risk factors for Lyme disease.
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Lyme Testing: Accurate When Used Appropriately
A 2016 systematic review that included 8 studies of CDC-recommended two tier test performance in Late Lyme showed Lyme antibody testing to be 99.4% sensitive and 99.3% specific** in North America.
In other words, out of 100 people with Late Lyme disease, 99-100 of them will test positive. Out of 100 people who may believe they have Late Lyme disease but do not, 99-100 of them will test negative.
Misinformation can convince patients to ignore or misinterpret negative tests to justify a false Chronic Lyme diagnosis.
How It Is Done
The health professional taking a sample of your blood will:
- Wrap an elastic band around your upper arm to stop the flow of blood. This makes the veins below the band larger so it is easier to put a needle into the vein.
- Clean the needle site with alcohol.
- Put the needle into the vein. More than one needle stick may be needed.
- Attach a tube to the needle to fill it with blood.
- Remove the band from your arm when enough blood is collected.
- Put a gauze pad or cotton ball over the needle site as the needle is removed.
- Put pressure on the site and then put on a bandage.
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What Is A Lyme Disease Blood Test
A Lyme disease blood test is used to determine if you have contracted Borrelia burgdorferi , the bacterium that causes Lyme disease. Lyme disease tests are conducted with a routine blood draw.
While there are other species of Borrelia that cause Lyme disease, B. burgdorferi is the most common cause in the United States. Most antibody tests in the United States only test for B. burgdorferi, but other species-specific tests are available depending on a persons travel history.
Lyme disease is transmitted to humans through ticks that are infected with Borrelia.
- dizziness
- shortness of breath
Lyme disease can be difficult to diagnose. Ticks are very small, and the bites are not always noticeable. Symptoms of the disease can vary from person to person. Not everyone experiences the classic bulls-eye rash pattern around a tick bite.
It should be noted that testing is not always required to make a diagnosis. For people with a classic bulls-eye rash living in a high risk area, testing is not recommended for diagnosis.
Your doctor will use the results of a Lyme disease antibody test, along with the report of your symptoms, to confirm a diagnosis.
Antibodies are proteins your body makes in response to foreign or harmful substances called antigens. Common antigens include:
- bacteria
- fungi
- chemicals
If you have never been exposed to B. burgdorferi, you will not have any Lyme disease antibodies in your bloodstream. In this case, your test will be negative.
Severity Of Lyme Disease
Misconception: B. burgdorferi infection is potentially lethal.
Science: Although Lyme disease can cause heart or brain abnormalities, there have been remarkably fewif anydeaths attributable to this infection.
LymeScience note: After this paper was published, the CDC three case studies of deaths associated with Lyme carditis, though two patients had preexisting heart conditions. While its not entirely clear if the infection caused the deaths, the CDC still reiterates, Prompt recognition and early, appropriate therapy for Lyme disease is essential.
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What Happens During Lyme Disease Testing
Lyme disease testing is usually done with your blood or cerebrospinal fluid.
For a Lyme disease blood test:
- A health care professional will take a blood sample from a vein in your arm, using a small needle. After the needle is inserted, a small amount of blood will be collected into a test tube or vial. You may feel a little sting when the needle goes in or out. This usually takes less than five minutes.
If you have symptoms of Lyme disease affecting your nervous system, such as neck stiffness and numbness in hands or feet, you may need a test of cerebrospinal fluid . CSF is a clear liquid found in your brain and spinal cord. During this test, your CSF will be collected through a procedure called a lumbar puncture, also known as a spinal tap. During the procedure:
References Choosing A Test
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Lyme Disease Test Western Blot
Labs performing a Western blot use electricity to separate proteins called antigens into bands. The read-out from the Western blot looks like a bar code. The lab compares the pattern produced by running the test with your blood to a template pattern representing known cases of Lyme disease. If your blot has bands in the right places, and the right number of bands, it is positive.
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. Different laboratories use different methods and criteria for interpreting the test, so you can have a positive test result from one lab and a negative test result from another.
For a comprehensive explanation of the western blot test, download Understanding Western Blot Lyme disease test.
The chart below will help you understand how to interpret the western blot test.
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:
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