A fluorescent bead-based multiplex assay for the simultaneous detection of antibodies to B. burgdorferi outer surface proteins in canine serum

Wagner, Bettina; Freer, Heather; Rollins, Alicia; Erb, Hollis N.

doi:10.1016/j.vetimm.2010.12.003

Cited by 36 publications

(47 citation statements)

References 44 publications

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“…The canine patient sera were submitted for serological Lyme testing to the Animal Health Diagnostic Center at Cornell University between July 2008 and January 2009, and antibodies to B. burgdorferi were analyzed by Western blotting using a whole-cell lysate from cultured bacteria (2,11). Whole-cell lysates from B. burgdorferi contain multiple antigens, including OspA, OspC, OspF, FlaB, and VlsE (39). Information about clinical signs or a disease history was not available for these dogs.…”

Section: Experimental Dogs and Samplesmentioning

confidence: 99%

“…burgdorferi antigens. A fluorescent bead-based multiplex assay was used to measure antibodies to B. burgdorferi OspA, OspC, and OspF antigens in canine serum (39). For this study, three additional antigens were added to the existing assay for the simultaneous measurement of antibodies to six antigens of B. burgdorferi.…”

Section: Experimental Dogs and Samplesmentioning

confidence: 99%

“…Fluorescent bead-based, multiplex analysis of antibodies to B. burgdorferi is a novel approach of high analytical sensitivity and allows for the simultaneous detection of immune responses to several antigens in dogs and horses (39,40). Several antigens of B. burgdorferi, including outer surface protein A (OspA), OspC, and OspF, are differentially expressed in ticks (OspA) (23,24,30,36,44) during transmission to a warm-blooded animal (OspC) (7,25,28) or later in the mammalian host (OspF) (1,18,21).…”

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confidence: 99%

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Antibodies to Borrelia burgdorferi OspA, OspC, OspF, and C6 Antigens as Markers for Early and Late Infection in Dogs

Wagner

Freer

Rollins

et al. 2012

Clin. Vaccine Immunol.

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ABSTRACTLyme disease in the United States is caused byBorrelia burgdorferisensu stricto, which is transmitted to mammals by infected ticks.Borreliaspirochetes differentially express immunogenic outer surface proteins (Osp). Our aim was to evaluate antibody responses to Osp antigens to aid the diagnosis of early infection and the management of Lyme disease. We analyzed antibody responses during the first 3 months after the experimental infection of dogs using a novel multiplex assay. Results were compared to those obtained with two commercial assays detecting C6 antigen. Multiplex analysis identified antibodies to OspC and C6 as early as 3 weeks postinfection (p.i.) and those to OspF by 5 weeks p.i. Antibodies to C6 and OspF increased throughout the study, while antibodies to OspC peaked between 7 and 11 weeks p.i. and declined thereafter. A short-term antibody response to OspA was observed in 3/8 experimentally infected dogs on day 21 p.i. Quant C6 enzyme-linked immunosorbent assay (ELISA) results matched multiplex results during the first 7 weeks p.i.; however, antibody levels subsequently declined by up to 29%. Immune responses then were analyzed in sera from 125 client-owned dogs and revealed high agreement between antibodies to OspF and C6 as robust markers for infection. Results from canine patient sera supported that OspC is an early infection marker and antibodies to OspC decline over time. The onset and decline of antibody responses toB. burgdorferiOsp antigens and C6 reflect their differential expression during infection. They provide valuable tools to determine the stage of infection, treatment outcomes, and vaccination status in dogs.

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Section: Experimental Dogs and Samplesmentioning

confidence: 99%

Section: Experimental Dogs and Samplesmentioning

confidence: 99%

mentioning

confidence: 99%

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Antibodies to Borrelia burgdorferi OspA, OspC, OspF, and C6 Antigens as Markers for Early and Late Infection in Dogs

Wagner

Freer

Rollins

et al. 2012

Clin. Vaccine Immunol.

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“…57 Lyme disease diagnostics have been developed for horses, dogs, and deer based on detection of antibodies to Borrelia burgdorferi OspA, OspC, and OspF antigens as indicators for infection. 78,[149][150][151][152] The assay distinguishes vaccination from infection (i.e., DIVA responses) and determines earlyversus late-infection stages to support treatment decisions. Such tests have also enabled researchers to use these host species as sentinels of Lyme disease ticks and diagnose Borrelia-associated uveitis in horses.…”

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confidence: 99%

Opportunities for bead-based multiplex assays in veterinary diagnostic laboratories

et al. 2013

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Abstract. Bead-based multiplex assays (BBMAs) are applicable for high throughput, simultaneous detection of multiple analytes in solution (from several to 50-500 analytes within a single, small sample volume). Currently, few assays are commercially available for veterinary applications, but they are available to identify and measure various cytokines, growth factors and their receptors, inflammatory proteins, kinases and inhibitors, neurobiology proteins, and pathogens and antibodies in human beings, nonhuman primates, and rodent species. In veterinary medicine, various nucleic acid and protein-coupled beads can be used in, or for the development of, antigen and antibody BBMAs, with the advantage that more data can be collected using approximately the same amount of labor as used for other antigen and antibody assays. Veterinary-related BBMAs could be used for detection of pathogens, genotyping, measurement of hormone levels, and in disease surveillance and vaccine assessment. It will be important to evaluate whether BBMAs are "fit for purpose," how costs and efficiencies compare between assays, which assays are published or commercially available for specific veterinary applications, and what procedures are involved in the development of the assays. It is expected that many veterinary-related BBMAs will be published and/or become commercially available in the next few years. The current review summarizes the BBMA technology and some of the currently available BBMAs developed for veterinary settings. Some of the human diagnostic BBMAs are also described, providing an example of possible templates for future development of new veterinary-related BBMAs.

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“…17 Detection of antibodies to OspA in the WB and bead assays has previously been associated with response to commercially available vaccines. 10,17 However, it was then documented that antibodies to OspA may occur in infected nonvaccinated dogs but the response is transient and lower than noted in vaccinated dogs. 18 Additionally, a commercially available vaccine a induces antibodies to both OspA and OspC, complicating differentiation of natural exposure and vaccinal response in the first few weeks of natural infection or a few weeks postvaccination.…”

Section: Introductionmentioning

confidence: 99%

Multiple antigen target approach using the Accuplex4 BioCD system to detect Borrelia burgdorferi antibodies in experimentally infected and vaccinated dogs

et al. 2015

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Abstract. The primary objective of our study was to optimize detection of serum antibodies to Borrelia burgdorferi using a new commercial automated fluorescence system (Accuplex4 BioCD system, Antech Diagnostics, Lake Success, New York). The system used multiple natural and artificial peptides-outer surface proteins (OspA, OspC, OspF), an outer membrane protein (P39), and a proprietary synthetic peptide (small Lyme peptide [SLP])-and the results were compared with a commercially available enzyme-linked immunosorbent assay that uses a proprietary peptide (C6). Sera from 4 groups were evaluated: dogs vaccinated with 1 of 3 commercially available vaccines (n = 18); dogs infested with adult Ixodes scapularis (black-legged tick; n = 18); dogs previously vaccinated and then infested with I. scapularis (n = 18); and dogs with B. burgdorferi infection that were then vaccinated (n = 14). All of the vaccines evaluated induced OspA responses. However, antibodies against OspF or C6 were not induced in any of the vaccinated dogs. Additionally, the OspF antibodies had 100% sensitivity and specificity when compared to antibodies against C6 peptide. In B. burgdorferi-infected dogs, antibodies against OspC and SLP were detected in serum sooner than antibodies against the other targets. Low levels of antibodies against OspA developed in 6 of 14 B. burgdorferi-infected, unvaccinated dogs and had the shortest duration compared to the other antibodies. Detection of antibody responses to multiple B. burgdorferi targets with this system can be used to help differentiate vaccinated dogs from exposed dogs as well as acute infection from chronic infection.

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A fluorescent bead-based multiplex assay for the simultaneous detection of antibodies to B. burgdorferi outer surface proteins in canine serum

Cited by 36 publications

References 44 publications

Antibodies to Borrelia burgdorferi OspA, OspC, OspF, and C6 Antigens as Markers for Early and Late Infection in Dogs

Antibodies to Borrelia burgdorferi OspA, OspC, OspF, and C6 Antigens as Markers for Early and Late Infection in Dogs

Opportunities for bead-based multiplex assays in veterinary diagnostic laboratories

Multiple antigen target approach using the Accuplex4 BioCD system to detect Borrelia burgdorferi antibodies in experimentally infected and vaccinated dogs

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