Ability of the Borreliacidal Antibody Test To Confirm Lyme Disease in Clinical Practice

Abstract

Highly specific borreliacidal antibodies are induced by infection with Borrelia burgdorferi, and a borreliacidal antibody test (BAT) may be an accurate laboratory procedure for confirming Lyme disease in clinical practice. To investigate this, 34 Lyme disease sera and 34 sera from patients with other illnesses who had presented to a primary-care facility located in an area of borreliosis endemicity were tested by the BAT and Western blotting (WB). The BAT was more sensitive (79% versus 65%; P ‫؍‬ 0.090), especially in cases in which patients had a single erythema migrans lesion (P ‫؍‬ 0.021). In addition, the potentially cross-reactive sera were negative by the BAT but WB yielded three (9%) false-positive results. The results from 104 sera from possible Lyme disease patients demonstrated the clinical usefulness of the more sensitive and specific BAT. The BAT was positive for 40 (38%) sera from patients with Lyme disease-related symptoms and appropriate clinical and epidemiological findings. WB confirmed Lyme disease in 30 (75%) of the 40 BAT-positive patients but failed to detect B. burgdorferi infection in 10 BAT-positive patients. WB was also positive for 11 BAT-negative sera, but six (55%) patients had case histories which suggested that the results were false positives. Collectively, the results confirm that the BAT is a sensitive and highly specific test and suggest that widespread use would increase the accuracy of serodiagnostic confirmation of Lyme disease.Lyme disease occurs after the bite of Borrelia burgdorferiinfected Ixodes spp. ticks. Detecting anti-B. burgdorferi antibodies in serum is the most commonly used method for confirming Lyme disease, since detecting the spirochetes by culture or PCR is problematic. The Centers for Disease Control and Prevention (CDC) recommends confirming Lyme disease by a two-tiered system, where serum is screened with an indirect immunofluorescence assay (IFA) or indirect enzymelinked immunosorbent assay. The serum with equivocal or positive results is then confirmed by immunoglobulin M (IgM) and IgG Western blotting (WB) or IgG WB alone if the illness has been present for less than or more than 1 month, respectively (14).WB can accurately confirm Lyme disease (21, 34, 35), especially when patients are tested who reside in a focus of endemicity and present with a tick exposure and well-recognized (13) clinical symptoms. However, false-positive WBs can occur (5, 15, 36) and the impact of the nonspecificity is magnified when the patients have less-typical symptoms or little risk of obtaining a tick bite (26,29,33,35). This shortcoming necessitates the development of a more accurate serodiagnostic test.Infection with B. burgdorferi induces the production of borreliacidal antibodies (6, 9-12, 17, 19, 24, 25, 30-32) that activate complement to form a membrane attack complex. The membrane attack complex then kills the spirochetes without the need for phagocytic cells (4,10,12,(22)(23)(24). Most B. burgdorferi proteins expressed on the surface of the spirochetes induce borrel...

Diagnosis of Lyme Borreliosis

“…Invariably these tests are nonreactive in healthy control populations or in sera of non-Lyme disease patients with rheumatological diseases (47,48). Modifications of the assay with reported higher sensitivities employ B. burgdorferi sensu stricto strain 50772, which lacks OspA and OspB (44,45). Major disadvantages include the need for cultured live B. burgdorferi sensu stricto, interference from antimicrobials that might be present in patient sera, and the relatively cumbersome nature of the assays.…”

One-Year Duration of Immunity Induced by Vaccination with a Canine Lyme Disease Bacterin

“…The borreliacidal antibodies were detected indirectly by monitoring the increased fluorescence intensity that occurs when the acridine orange intercalates into blebbed, nonviable spirochetes. A Ն13% shift in the mean fluorescence intensity compared to a normal serum control was considered positive (2). The presence of blebbed nonmotile B. burgdorferi was confirmed by dark-field microscopy.…”

“…Blood samples were obtained 7 days (day 28) after the booster and immediately prior (day 394) to the tick challenge and tested for anti-OspA or anti-OspC borreliacidal antibodies as described previously (2). Briefly, 5 ϫ 10 5 lowpassage B. burgdorferi S-1-10 (OspA) or 50772 (OspC) organisms were combined with serum and guinea pig complement (Rockland Immunochemical, Gilbertsville, PA), and the suspension was incubated at 35°C.…”

Vaccination with theospA- andospB-Negative Borrelia burgdorferi Strain 50772 Provides Significant Protection against Canine Lyme Disease

“…The borreliacidal antibodies were detected indirectly by monitoring the increased fluorescence intensity that occurs when the acridine orange intercalates into blebbed, nonviable spirochetes. A Ն13% shift in the mean fluorescence intensity compared to a normal serum control was considered positive (21). The presence of blebbed, nonmotile B. burgdorferi was confirmed by dark-field microscopy.…”

“…Blood samples were collected immediately prior to the tick challenge (7 days after booster) and tested for anti-OspA or anti-OspC borreliacidal antibodies, as described previously (17,21). Briefly, 5 ϫ 10 5 low-passage-number B. burgdorferi strains S-1-10 (OspA) or 50772 (OspC) were combined with serum and guinea pig complement, and the suspension was incubated at 35°C for 16 to 24 h. Following incubation, 100 l of each assay suspension was combined with phosphate-buffered saline (PBS) and acridine orange, and the spirochetes were monitored for killing by using a FACScan flow cytometer (Becton Dickinson Immunocytometry Systems, San Jose, CA).…”