Detection of Borreliacidal Antibodies by Flow Cytometry

Callister, Steven M.; Schell, Ronald F.; Lim, L. C. L.; Jobe, Dean A.; Case, Kay L.; Bryant, Gary L.; Molling, Paul

doi:10.1001/archinte.1994.00420140095012

Cited by 31 publications

(10 citation statements)

References 41 publications

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“…As seen in our experimental dogs, the levels of growth-inhibiting antibodies were low or the antibodies were missing after exposure to ticks. Eventually, a more sensitive test based on flow cytometry used by Callister et al (6) may provide a helpful tool for the diagnosis of early Lyme disease. At this time it is not clear whether this method will be applicable during the early stage of the disease, since it is not known when borreliacidal antibodies appear in detectable quantities.…”

Section: Discussionmentioning

confidence: 99%

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Sera from OspA-vaccinated dogs, but not those from tick-infected dogs, inhibit in vitro growth of Borrelia burgdorferi

et al. 1995

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Dogs were challenged with Borrelia burgdorferi by exposure to ticks, with or without prior protection from infection by recombinant OspA (rOspA) vaccination. Sera from these dogs were tested for their capability to inhibit the growth of B. burgdorferi in vitro. Bacterial growth was detected by a color change in the culture medium, and the optical density was measured with a spectrophotometer in microtiter plates. By growth inhibition, which was complement dependent, the color change was lacking after 5 days of incubation. Over a 1-year study, nonvaccinated dogs infected by exposure to ticks showed high antibody titers to B. burgdorferi by kinetic enzyme-linked immunosorbent assay (KELA). The same sera did not inhibit spirochetal growth or did so only at a low dilution. These results corresponded to the lack of OspA and OspB antibodies seen in Western blots (immunoblots), and these dogs were not protected from infection or disease. In contrast, dogs immunized with rOspA prior to challenge with infected ticks produced high antibody titers, as determined by KELA, but their sera also had high growth-inhibiting antibody titers. Western blot analysis showed a strong band in the 32-kDa region when the sera of these dogs were tested. When adjuvant was administered with rOspA, antibody titers by both KELA and growth inhibition were higher and persisted longer in the immunized dogs. All dogs immunized with rOspA were protected from infection and disease.

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Section: Discussionmentioning

confidence: 99%

“…The color change in the culture medium induced by live spirochetes was found to be a reliable and less labor intensive method of measuring growth-inhibiting antibodies (26,36,37). Recently, it was shown that flow cytometry is a useful tool for identifying infected patients apparently with small quantities of borreliacidal antibodies (6).…”

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confidence: 99%

Sera from OspA-vaccinated dogs, but not those from tick-infected dogs, inhibit in vitro growth of Borrelia burgdorferi

et al. 1995

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“…Detection of borreliacidal antibodies. Borreliacidal antibodies were detected with a flow cytometer as previously described (7,10,22). Briefly, a frozen aliquot of B. burgdorferi sensu stricto isolate 297 or 50772 was thawed, 200 l was inoculated into 6 ml of fresh BSK medium, and the cultures were incubated at 35°C for 72 h. Following incubation, the concentration of B. burgdorferi organisms was adjusted to 10 6 spirochetes per ml with fresh BSK medium.…”

Section: Methodsmentioning

confidence: 99%

Detection of Borreliacidal Antibodies in Lyme Borreliosis Patient Sera Containing Antimicrobial Agents

Jobe

Rawal

Schell

et al. 1999

Clin Diagn Lab Immunol

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The borreliacidal-antibody test has been used for the serological detection and confirmation of Lyme borreliosis. However, the presence of antimicrobial agents in serum can confound the accurate detection of borreliacidal antibodies. In this study, we developed a Bacillus subtilis agar diffusion bioassay to detect small concentrations of antimicrobial agents in serum. We also used XAD-16, a nonionic polymeric resin, to adsorb and remove high concentrations of amoxicillin, cefotaxime, ceftriaxone, cefuroxime, doxycycline, and erythromycin without significantly affecting even small concentrations of immunoglobulin M (IgM) or IgG borreliacidal antibodies. High concentrations of penicillin could also be removed by adding 1 U of penicillinase without significantly influencing the levels of borreliacidal antibodies. These simple procedures greatly enhance the clinical utility of the borreliacidal-antibody test.

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“…More recently recombinant chimeric proteins have been created, combining critical epitopes from several immunodominant antigens [23]. Another experimental assay uses flow cytometry to detect borreliacidal antibodies which may be more clinically relevant [24]. However, a recent study found quite low sensitivity (11%) for this assay in active Lyme disease [14].…”

Section: Advancesmentioning

confidence: 99%

Advances and pitfalls in the diagnosis of Lyme disease

Coyle

2006

FEMS Immunology &Amp; Medical Microbiology

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Detection of Borreliacidal Antibodies by Flow Cytometry

Abstract: Detection of borreliacidal antibody, unlike indirect fluorescent antibody and enzyme immunoassay, is an accurate, highly specific serodiagnostic test for detection of Lyme disease.

Cited by 31 publications

References 41 publications

Sera from OspA-vaccinated dogs, but not those from tick-infected dogs, inhibit in vitro growth of Borrelia burgdorferi

Sera from OspA-vaccinated dogs, but not those from tick-infected dogs, inhibit in vitro growth of Borrelia burgdorferi

Detection of Borreliacidal Antibodies in Lyme Borreliosis Patient Sera Containing Antimicrobial Agents

Advances and pitfalls in the diagnosis of Lyme disease

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