Texas A& m University Research Aims To Improve Lyme Disease Diagnostics
Story by Rachel Knight, VMBS Communications
Lyme disease, the fastest growing vector-borne illness in the U.S., according to the Bay Area Lyme Foundation, is challenging to diagnose and can only be treated in the early stages of infection. Once the infection spreads to the nervous and muscular systems, it is both harder to detect and less susceptible to antibiotics.
Research by two Texas A& M University scientists, however, is focused on improving Lyme disease treatment outcomes by developing a test thats both more accurate and more efficient than the current test for the infection.
Dr. Artem Rogovskyy, an associate professor at the Texas A& M School of Veterinary Medicine & Biomedical Sciences , and Dr. Dmitry Kurouski, an assistant professor in the Texas A& M Department of Biochemistry & Biophysics and the Department of Biomedical Engineering, are testing Raman spectroscopy, a technique used to detect vibrations at the molecular level, as a diagnostic tool for Lyme disease.
The results of Rogovskyy and Kurouskis second paper on Raman spectroscopy as a diagnostic tool for Lyme disease, published in October, demonstrate that blood samples from mice and humans infected with the Lyme pathogen were more accurately identified with the Raman spectroscopy test than with the two-tiered serology, the only diagnostic method currently approved to diagnose Lyme disease in humans in the United States.
You Do Not Usually Need Tests To Show That You Have Lyme Disease
In most cases, theres a clear sign of Lyme diseasea painless, spreading rash that often grows to look like a bulls eye. If you have this rash, and you recently had a tick bite or were in an area known for Lyme disease, you dont need a test. Instead, your doctor can just start treating you with antibiotics, as appropriate.
Interpretation Of Lyme Multiplex Results1213
The Lyme Multiplex Assay is a fully quantitative test. It results in a numeric antibody value for each of the three B. burgdorferi antigens tested. A brief interpretation on each value is submitted with the test report. In addition, the antibody profile gives an advanced interpretation on the infection and vaccination status of the horse. Antibodies to OspA serve as markers for vaccination and those to OspC and OspF as markers for infection . In treated horses, quantitative antibody values are valuable indicators to follow-up on treatment success.
Figure 3:B. burgdorferi
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Important: Dont Misinterpret A Negative Test As Positive
Many people without Lyme disease will test positive for some bands. Therefore, CDC cautions:
It is not correct to interpret a test result that has only some bands that are positive as being mildly or somewhat positive for Lyme disease.
For example, in one study, 43% of healthy people and 75% of syphilis patients tested positive for IgG band 41. In a study of US veterans in New York, 76% of those without Lyme disease tested positive for IgG band 41. In a 1996 study, in healthy people, 55% and 21% tested positive for IgG band 41 and IgM band 41, respectively.
Even without a Borrelia burgdorferi infection, many of us produce antibodies that will react on a Lyme test. Notably, harmless bacteria found naturally in our mouths can cause us to test positive for band 41.
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Who Should Get Testing
Testing is usually indicated if a person has symptoms of Lyme disease and a known or possible exposure to ticks that can carry the Borrelia bacteria. However, because it takes time for antibodies to develop, the timing of testing is important to consider.
Lyme disease symptoms depend on the extent of the bacterial infection. Three phases are used to describe the infection:
It is important to understand these phases because testing is not equally valuable in each stage. With early localized disease and erythema migrans, blood testing is generally not helpful because antibodies have not had enough time to develop.
Because of similar concerns about potential false positive results, random screening for Lyme disease in people without symptoms is not recommended even in areas that are known to have ticks that can carry the Borrelia bacteria.
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Lyme Disease In Europe And Asia
Lyme disease can also occur in Europe and Asia, where Borrelia garinii and Borrelia afzelii are most commonly found.
Ticks infected with a Lyme disease bacterium can be found in woodlands across the European continent from northern Turkey to northern Sweden. However, Lyme disease is considered endemic in central Europe, where the following countries have the highest tick infection rates: Austria, Czech Republic, southern Germany,Switzerland, Slovakia, and Slovenia. In Europe, Lyme disease is primarily transmitted by the castor bean tick.
Lyme disease has been reported throughout Asia, as well, such as in Russia, Mongolia, northern China, Japan, and Koreaâ âthough infection from a Lyme disease bacterium appears to be relatively uncommon in these areas. In Asia, Lyme disease is transmitted by the taiga tick .
Special Considerations For Vaccinated Horses
An approved Lyme vaccine for horses is currently not available. Horses are sometimes vaccinated with one of the three available Lyme vaccines for dogs for attempted protection of horses that are housed in Lyme endemic areas. Efficacy studies of canine vaccines in horses are not yet available, but experimental data suggested that anti-OspA antibodies are protective in horses.
All available vaccines contain OspA antigen as the sole or one of the vaccine components. Antibodies to OspA are identified by the Equine Lyme Multiplex Assay to determine the vaccination status in vaccinated horses. To provide our clients with the best interpretation for each animal, we need information on the vaccine used. This includes the name of the vaccine and the date when the horse was last vaccinated. Please include this information on the submission form when samples of vaccinated horses are submitted for testing.
Recent studies indicate that horses can respond to the canine vaccines, but the responses are typically short-lived17,18. It is therefore recommended to vaccinate horses in close proximity to tick season, approximately four weeks before ticks are typically abundant in the area. Additionally, confirmatory testing is recommended after vaccination of horses since low OspA antibody responses have been observed in individual horses.
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Serologic Tests Are The Gold Standard
Prompt diagnosis is important, as early Lyme disease is easily treatable without any future sequelae.
Tests for Lyme disease can be divided into direct methods, which detect the spirochete itself by culture or by polymerase chain reaction , and indirect methods, which detect antibodies . Direct tests lack sensitivity for Lyme disease hence, serologic tests remain the gold standard. Currently recommended is a standard 2-tier testing strategy using an enzyme-linked immunosorbent assay followed by Western blot for confirmation.
Diagnostic testing methods in Lyme disease
Where Is Lyme Disease Found
In the United States, Lyme disease has been reported in every state, but over 95% of cases are from the Northeastern, Mid-Atlantic, and upper Midwestern states, with a small number of cases reported along the West Coast, especially Northern California. In Canada, Lyme-positive dogs are found mostly in southern Ontario and southern Manitoba, with a small number of cases in southern Quebec and the Maritime provinces.
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How Does The Equine Lyme Multiplex Assay Work
The Equine Lyme Multiplex Assay was developed at the Animal Health Diagnostic Center at Cornell University. It detects antibodies to three antigens of B. burgdorferi in equine serum . The test is based on fluorescent beads and allows the simultaneous measurement of antibodies to all three B. burgdorferi antigens in a single sample13.
Figure 2:B. burgdorferi
Lyme Igm Antibody Serology
The IgG/IgM Antibody Serology test is an ELISA format, and it detects the presence of IgM antibodies to B. burgdorferi after exposure to an infected tick. Because IgM antibodies appear early in response to infection, this test may be positive two to six weeks after exposure. But since the level of IgM rapidly declines over time, testing for IgM antibodies too late can cause a missed infection.
A positive or equivocal IgM antibody test must be confirmed by an IgM Western Blot or Lyme ImmunoBlot IgM. NOTE: The sensitivity concerns mentioned for the IgG/IgM assay also affect this assay.
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Is There A Lyme Disease Test Kit
Blood collection kits are available for at-home Lyme disease testing. Lyme disease test kits can cost as less as $20 and as much as $100 or more. Using a Lyme disease test kit is as simple as pricking your finger and smearing or collecting the blood onto the kit for testing.
However, testing for Lyme disease in a more controlled environment such as a lab or clinic is preferable as qualified healthcare professionals are likely to perform a more reliable test.
Waddell L. A., et al. . The accuracy of diagnostic tests for Lyme disease in humans, a systematic review and meta-analysis of North American research. PLoS One. 2016 11: e0168613. Retrieved from
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.
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Negative Elisa On Serum
Early clinical Lyme disease in the form of erythema migrans with an associated history of a tick bite should be treated empirically. There is no need for testing unless there are further symptoms.
A negative ELISA result in the early stages of Lyme disease does not exclude infection. If acute Lyme disease is suspected but serology results are negative, we recommend that the test is repeated in 4 to 6 weeks with a fresh sample to look for seroconversion.
In patients with long term symptoms a negative ELISA test usually excludes Lyme disease as a cause of these symptoms. Information on differential diagnosis for patients with persistent symptoms and negative Lyme disease serology results is available.
Lyme Disease Laboratory Tests Available At Ripl
The primary service provided by RIPL is serological testing using well-characterised and validated screening and confirmatory tests in accordance with the NICE Lyme disease guideline.
RIPL participates in regular external quality assurance exercises as an independent measure of its performance.
Details of prices and turnaround times for Lyme testing are provided in Appendix 1 of the RIPL user manual.
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Diagnosis Testing And Treatment
You may have heard that the blood test for Lyme disease is correctly positive only 65% of the time or less. This is misleading information. As with serologic tests for other infectious diseases, the accuracy of the test depends upon how long youve been infected. During the first few weeks of infection, such as when a patient has an erythema migrans rash, the test is expected to be negative.
Several weeks after infection, FDA cleared tests have very good sensitivity.
It is possible for someone who was infected with Lyme disease to test negative because:
If you are pregnant and suspect you have contracted Lyme disease, contact your physician immediately.
* Silver HM. Lyme disease during pregnancy. Infect Dis Clin North Am. 1997 Mar 11:93-7.
The most common co-infections that occur with Lyme disease are anaplasmosis and babesiosis. In general:
When Not To Take This Lyme Disease Test
Do not take this Lyme disease antibody test if:
- You are experiencing a round rash after a tick bite, such as the typical bullseye rash associated with early Lyme disease. Seek immediate medical attention from a health professional instead of taking this test. A round rash could be a sign of Lyme disease, and it’s best to consult a medical professional and receive Lyme disease treatment as soon as possible to help prevent complications associated with the disease.
- You suspect you might have been infected with Lyme disease bacteria less than 6 weeks ago . The Lyme disease antibodies detected by this test take several weeks to build up in your bloodstream, taking the test before 6 weeks have passed may result in a false negative.
- You havenât lived in or traveled to an area where Lyme disease occurs. Consider discussing any symptoms with your healthcare providerâinstead of taking this testâto help determine if youâve been exposed to a Lyme disease bacterium.
- Youâve previously received a Lyme disease diagnosis, was treated for the disease, and are still symptomatic. Consider discussing any new or ongoing symptoms with your healthcare provider instead of taking this test.
- You are currently being treated for Lyme disease infection or taking antibiotic treatment for other infections. Consider discussing any symptoms with your healthcare provider instead of taking this test.
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What Happens During Lyme Disease Testing
Lyme disease testing is usually done on a sample of blood. In certain cases, a cerebrospinal fluid test may be done.
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.
For a CSF test:You may need a cerebrospinal fluid test if your symptoms could mean that Lyme disease is affecting your nervous system, such as a stiff neck or numb hands or feet. Providers may order a CSF test if the results of your blood test show you likely have Lyme disease or if the results are uncertain.
To get a sample of your cerebrospinal fluid, a provider will do a procedure called a lumbar puncture, also known as a spinal tap. During the procedure:
Laboratory Tests That Are Not Recommended
Some laboratories offer Lyme disease testing using assays whose accuracy and clinical usefulness have not been adequately established. Examples of unvalidated tests include:
- Capture assays for antigens in urine
- Culture, immunofluorescence staining, or cell sorting of cell wall-deficient or cystic forms of B. burgdorferi
- Lymphocyte transformation tests
- In-house criteria for interpretation of immunoblots
- Measurements of antibodies in joint fluid
- IgM or IgG tests without a previous ELISA/EIA/IFA
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Why Do I Need A Lyme Disease Test
You may need a test if you have symptoms of infection and were exposed or possibly exposed to ticks that carry the bacteria that causes Lyme disease. The first symptoms of Lyme disease usually show up between 3 and 30 days after the tick bite.
Early signs and symptoms may include:
- A reddish round skin rash that slowly gets bigger over several days. It usually doesn’t itch or hurt, but it may be warm. As it gets better, it may look like a bull’s-eye , but not always.
- Muscle and joint aches
- Swollen lymph nodes
Later signs and symptoms of Lyme disease usually affect your joints and nervous system. They include:
- Severe headaches and neck stiffness
- Rashes on other parts of your body
- Arthritis with severe joint pain and swelling, especially in the knees
- Pain that comes and goes in your muscles, joints, bones, and tendons
- Shooting pains, numbness, or tingling in your hands or feet
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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