Serologic Tests For Lyme Disease Research Articles

Molecular characterization of the humoral response to the 41-kilodalton flagellar antigen of Borrelia burgdorferi, the Lyme disease agent

The earliest humoral response in patients infected with Borrelia burgdorferi, the agent of Lyme disease, is directed against the spirochete's 41-kDa flagellar antigen. In order to map the epitopes recognized on this antigen, 11 overlapping fragments spanning the flagellin gene were cloned by polymerase chain reaction and inserted into an Escherichia coli expression vector which directed their expression as fusion proteins containing glutathione S-transferase at the N terminus and a flagellin fragment at the C terminus. Affinity-purified fusion proteins were assayed for reactivity on Western blots (immunoblots) with sera from patients with late-stage Lyme disease. The same immunodominant domain was bound by sera from 17 of 18 patients. This domain (comprising amino acids 197 to 241) does not share significant homology with other bacterial flagellins and therefore may be useful in serological testing for Lyme disease.

Problems in the Use of Serologic Tests for the Diagnosis of Lyme Disease

Lyme disease can be reliably diagnosed in the presence of erythema migrans. When erythema migrans is absent, serologic tests are often used to confirm the diagnosis. To choose a test for our Lyme disease diagnostic center, serum samples were obtained from 34 patients and tested for antibodies to Borrelia burgdorferi. We evaluated five enzyme-linked immunosorbent assays from Stony Brook (NY) University Hospital, Cambridge Bioscience (Worcester, Mass), Hillcrest Biologicals (Cypress, Calif), Sigma Diagnostics (St Louis, Mo), and Zeus-Wampole Scientific Inc (Raritan, NJ) and two fluorescent antibody tests (3M [Diagnostic Systems Inc, Santa Clara, Calif] and FIAX [Whittaker M.A. Bioproducts Inc, Walkersville, Md]). A positive sample by any test was further analyzed by Western blot. Using the Centers for Disease Control (Atlanta, Ga) epidemiologic case definitions, patients were classified into those with clinical Lyme disease, patients not meeting the Centers for Disease Control definitions, and asymptomatic patients. Sensitivities of Lyme serologies varied from 13% to 73%, with Hillcrest showing the highest value and Sigma the lowest value. False-positive test results were found in 0% to 27% of patients. Western blot analysis was positive in six of 15 patients with clinical Lyme disease. These results emphasize the need for better serologic testing for Lyme disease and underline their usefulness only as adjuncts in the clinical diagnosis of Lyme disease.

Serodiagnosis of Lyme Disease

Letters15 June 1991Serodiagnosis of Lyme DiseaseJeffrey M. Jones, MD, PhDJeffrey M. Jones, MD, PhDSearch for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-114-12-1064_1 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptTo the Editors: Rahn and Malawista (1) rightly point out that serologic testing for Lyme disease is poorly standardized. In addition to the problem of interlaboratory reproducibility of results, another problem must be considered: None of the currently used antibody tests has been rigorously proved to detect antibodies specific for Borrelia burgdorferi. Too few control sera specimens have been studied to evaluate specificity adequately, given the widespread use of these tests.Wilske and colleagues (2) and Hunter and coworkers (3) studied 13 sera specimens with anti-B?. burgdorferi immunofluorescentantibody (IFA) titers of 1:256 or higher from patients with well-documented Lyme disease....

Frequency and clinical significance of Lyme seropositivity in patients with isolated optic neuritis

We evaluated antibody reactivity against Borrelia burgdorferi in 20 consecutive patients with newly diagnosed isolated optic neuritis who resided in a region endemic for Lyme disease. Four (20%) patients had positive serology. All three patients who had follow-up serologies showed rising convalescent levels of Borrelia-specific IgM. One patient refused lumbar puncture, one had normal CSF constituents except for an elevated Lyme antibody index, and two had CSF lymphocytic pleocytosis that remained unexplained after extensive evaluations for causes other than Lyme disease. We treated both patients who had CSF pleocytosis with intravenous ceftriaxone; the pleocytosis and optic nerve function improved. The other two patients received oral antibiotics and showed excellent recovery of visual acuity. We believe that serologic testing for Lyme disease is warranted for individuals with optic neuritis who reside in an endemic region, and patients with rising convalescent antibody levels or unexplained CSF pleocytosis should receive antibiotic treatment for Lyme disease.

Lyme Test Problems

<h3>To the Editor.—</h3> I suspect a methodological error in the recent article on serologic tests for Lyme disease by Luger and Krauss.¹The authors report that one of the university laboratories detected antibodies to<i>Borrelia burgdorferi</i>in 18 (100%) of the 18 specimens, while all other reference laboratories (n = 8) detected<i>B burgdorferi</i>in significantly fewer specimens (44% to 78%). The error likely resides in Luger and Krauss' use of the same university laboratory for inclusion into the study as well as for use as a reference laboratory. I would expect close to a 100% correlation using the same technique in the same laboratory. The laboratory used to meet the entrance criteria should have been excluded from the studied reference laboratories. If Luger and Krauss had excluded the laboratory used to meet the entrance criteria, then the study should have commented on the techniques used by laboratories having a

Cross-reactivity of nonspecific treponemal antibody in serologic tests for Lyme disease

Serum samples obtained from 59 persons who had acute necrotizing ulcerative gingivitis, periodontitis, syphilis, or Lyme disease were tested against Treponema phagedenis biotype Reiter, Treponema denticola, Treponema vincentii, and Treponema scoliodontum by indirect fluorescent-antibody staining methods. Although there were positive reactions for sera representing each of these study groups and for 20 (13%) of 156 samples collected from the general population (premarital screening for syphilis), titration endpoints were relatively low (less than or equal to 1:256). Serum samples from 18 persons who had gingivitis or periodontitis but no history of Lyme borreliosis were tested by enzyme-linked immunosorbent assay for antibodies to Borrelia burgdorferi. Of these, five (28%) had immunoglobulin M antibody and four (22%) contained immunoglobulin G antibodies to this spirochete. Adsorption with either sorbent commercially prepared from T. phagedenis biotype Reiter or with washed, whole cells of T. phagedenis biotype Reiter reduced cross-reactivity in the enzyme-linked immunosorbent assay for Lyme borreliosis.

Serologic testing for Lyme disease

Serologic testing for antibodies is, at present, the only practical means of diagnosing Borrelia burgdorferi infection. However, specific IgM spirochetal antibodies may not be at detectable concentrations in the early stage of Lyme disease (within 6 weeks of onset). Western blot analysis may be used to confirm indirect fluorescent antibody staining or enzyme-linked immunosorbent assay results, but it is laborious and expensive. False-positive reactions caused by cross-reactivity do occur. In patients who have had subclinical Lyme disease and contract another illness that causes similar symptoms, interpretation of serologic test results may be confused by residual antibodies. Therefore, assay results should always be interpreted in conjunction with clinical and epidemiologic findings.

Serologic Tests for Lyme Disease

The serologic test for the detection of antibodies to Borrelia burgdorferi is the most frequently used laboratory method for the diagnosis of Lyme disease. However, the insensitivity of the assays and the interlaboratory variability are frequent problems. To determine the extent of this variability, one aliquot of serum from each of nine patients with a history of Lyme disease was sent to nine reference laboratories, including national, university, state, and local hospital laboratories. A second aliquot of the original serum was submitted 2 weeks later. Wide variability among laboratories was observed, ranging from a university laboratory that detected antibody to B burgdorferi (IgG or IgM) in 18 of 18 specimens, to a state laboratory that detected antibody in only 8 of 18 specimens. Detection of IgM specific antibodies showed similar variability (range, 2 to 10 of 18). There were eight instances of a fourfold or greater change in titer between the aliquots sent 2 weeks apart, although only three of these were an increase in titer. These results indicate the need for standardization of the assays and the availability of national reference material. It is recommended that the results of serologic testing should not be relied on as the sole criteria in making the diagnosis of Lyme disease.

Serologic tests for Lyme disease. Interlaboratory variability

Serologic tests for Lyme disease. Interlaboratory variability