What is the recommended testing protocol for Lyme disease?

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From the Guidelines

The recommended testing protocol for Lyme disease is a two-tier approach, starting with an enzyme immunoassay (EIA) or immunofluorescence assay (IFA), followed by a Western blot test if the initial result is positive or equivocal, as recommended by the most recent guidelines 1. This approach is necessary to balance sensitivity and specificity, as the initial screening tests can yield false-positive results, while the Western blot test provides higher specificity to confirm the diagnosis.

  • The two-tier testing approach is particularly important for patients with symptoms consistent with Lyme disease who have been in endemic areas where the disease-carrying ticks are prevalent.
  • It's crucial to note that testing during the early stage of infection (within the first few weeks) may produce false-negative results since antibodies take time to develop, as highlighted in studies 1.
  • In cases where a patient presents with the characteristic erythema migrans rash and has a history of tick exposure in an endemic area, treatment can begin without waiting for test results.
  • For patients with later manifestations of Lyme disease, both IgM and IgG antibody tests are useful, though IgG becomes more relevant after the first few weeks of infection.
  • If neurological symptoms are present, cerebrospinal fluid testing may also be recommended to check for antibodies or inflammatory markers, as suggested by recent guidelines 1.

From the Research

Testing for Lyme Disease

The recommended testing protocol for Lyme disease involves a combination of laboratory tests to help diagnose Borrelia burgdorferi infections. Some of the key tests used include:

  • Indirect fluorescent antibody (IFA) staining methods
  • Enzyme-linked immunosorbent assay (ELISA)
  • Western blot analyses
  • Culturing or DNA detection by polymerase chain reaction (PCR) methods

Laboratory Diagnosis

According to 2, laboratory tests have been used extensively to help diagnose Borrelia burgdorferi infections. The results of indirect fluorescent antibody (IFA) staining methods or an ELISA, combined or separate from findings of Western blot analyses, have confirmed clinical diagnoses of Lyme disease.

Western Blotting

3 found that Western blotting is a useful adjunct in the serodiagnosis of Lyme neuroborreliosis and other manifestations of Lyme disease. Using a gel densitometric analysis, they devised quantitative criteria for positivity and tested their criteria by matching blot results with clinical characteristics in a retrospectively studied group of patients.

Two-Tier Assay

4 noted that the laboratory diagnosis of Lyme disease is based upon the detection of antibodies generated against Borrelia burgdorferi using a two-tier assay, typically consisting of an ELISA, followed by a Western blot. This system is insensitive for diagnosing early infection, when most people seek care.

Evaluation of Serological Tests

5 compared the performance of 11 commercially available enzyme immunoassays (EIA) and four Western blot (WB) tests for the detection of IgM and IgG antibodies against Borrelia burgdorferi. They found that the sensitivity of the individual tests ranged from 35 to 81% for detection of IgM in specimens from patients with early Lyme borreliosis.

Serologic Diagnosis

6 used class-specific enzyme-linked immunosorbent assays (ELISAs) with purified recombinant antigens of Borrelia burgdorferi sensu stricto and Western blot analyses with whole cells of this spirochete to test human sera. They found that there was frequent antibody reactivity to protein 41-G (p41-G), outer surface protein C (OspC), and OspF antigens in analyses for immunoglobulin M (IgM) antibodies.

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Research

Current status of laboratory diagnosis for Lyme disease.

The American journal of medicine, 1995

Research

Evaluation of fifteen commercially available serological tests for diagnosis of Lyme borreliosis.

European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, 1999

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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