Because the first test may not pick up antibodies soon after possible exposure to Lyme disease, that means a person with Lyme may never get the second test. The CDC testing guidelines recommend a second test only if the first test is positive or unclear. A possible drawback of the two-step testing system You should not need to go back to the lab. These tests can be run on the blood sample used for the first test. Because these tests are newer, not all doctors have experience with them and not all labs have access to them. The other option is to do a second ELISA test, which measures antibodies to a specific protein found on Lyme disease bacteria. The Western blot test has been part of the two-step testing system since 1994 and is more commonly used. One is called a Western blot, which looks at how your antibodies react to specific parts of the Lyme disease bacteria. There are two options for the second test. The second test: Western Blot or a second ELISA Because it can take some time for your body to produce antibodies, this test isn’t always accurate soon after a person is infected. This blood test is for antibodies against the Lyme disease bacteria. The first test: ELISA (enzyme-linked immunosorbent assay) Using today’s tests it is difficult to distinguish between current and previous infections, so if you were previously diagnosed with Lyme disease and may have been re-infected, your doctor should make the diagnosis based on your symptoms and risk factors. In some cases, a Lyme test may not be helpful. If you think you may have Lyme disease, but your doctor hasn’t considered it or has ruled it out before testing you for it, here’s how you can start a conversation: Give your doctor a list of Lyme disease symptoms and risk factors, and ask them if, given your own symptoms and risk factors, Lyme disease could be a possible cause and whether you should be tested. If you have been traveling, your doctor might test you for species of Borrelia that cause Lyme disease in other parts of the world. Your doctor may also do tests to see if other conditions with similar or overlapping symptoms - for example, thyroid disease, autoimmune disease, or other tick-borne diseases - could be causing your symptoms. Laboratory tests can support a clinical diagnosis, but should not be used on their own to confirm or rule out a current Lyme disease diagnosis. Your doctor will consider whether your symptoms and risk factors suggest Lyme disease when deciding whether you need a Lyme test. By that time, Lyme disease is harder to treat, and long-term complications are more likely, so an early, accurate diagnosis is very important. That’s why Lyme disease cases may be missed, and as many as 40% of Lyme disease cases are not diagnosed until a later stage of the disease. Using today’s tests, it is difficult to distinguish between an old Lyme disease infection and a new one. This may result in Lyme diagnoses being missed because symptoms are attributed to another illness. Symptoms of Lyme disease like fever and body aches can also be caused by other illnesses. It is also possible that a person may not notice a rash before it goes away. But up to 30% of people who get Lyme disease never get a rash. The only sure sign of Lyme disease is an erythema migrans (EM) rash, which often has a bullseye shape. That’s because Lyme tests look for proteins created by your immune system to help you fight off infection (antibodies), rather than for the infection itself, and it can take a few weeks for your body to make enough antibodies to cause a positive test. Tests are less accurate for a short period of time after you first become infected. Alfa Aesar offers a variety of reagents for Western blotting including enzyme substrates and blocking, transfer and washing buffers.There are several reasons it can be hard to diagnose Lyme disease. Depending on the substrates used, the conjugated enzymes can be detected by colorimetric, fluorescent or chemiluminescent techniques. With ELISA and Western blotting, the presence and/or amount of the antigen is determined directly or indirectly by an enzyme (usually horseradish peroxidase or alkaline phosphatase)-, biotin- or fluorophore-conjugated antibody. ELISA also allows quantification of the antigen in question. Like Western blots, ELISA is used to detect peptides in a mixture. Antibody binding to the target protein on the membrane can then be detected by various methods. A protein mixture is separated by polyacrylamide gel electrophoresis before transfer to a membrane. Western blotting is used to detect a specific target protein out of a complex protein mixture. Both techniques rely on immunodetection of an antibody to a specific protein. Western blotting and ELISA and are analytical techniques used to detect specific proteins in a biological sample.
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