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

Abstract

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.

ACVIM consensus update on Lyme borreliosis in dogs and cats

“…86 In other studies of dogs experimentally infected with Bb, anti-OspC antibodies were detected before those against other peptides, suggesting that the multiplex assay may be more sensitive in acute cases of LB. [83][84][85] months (without re-exposure), whereas OspF antibodies increase by 6-8 weeks and remain increased in untreated carriers. [83][84][85]87 The OspC antibodies probably increase again in field conditions (ie, re-exposure is a natural booster), thus finding OspC antibodies in a nonvaccinated dog may indicate recent exposure or re-exposure, without specifying when the dog was first infected in its life.…”

“…Vaccines that induce OspC antibodies interfere with this marker of natural exposure on Western blot, AccuPlex4, VetScan and Multiplex tests. After experimental natural exposure, OspC antibodies increase by 2–3 weeks, and wane in 3–5 months (without re‐exposure), whereas OspF antibodies increase by 6–8 weeks and remain increased in untreated carriers . The OspC antibodies probably increase again in field conditions (ie, re‐exposure is a natural booster), thus finding OspC antibodies in a nonvaccinated dog may indicate recent exposure or re‐exposure, without specifying when the dog was first infected in its life.…”

“…In dogs, serology is the only recommended modality to evaluate for exposure to Bb (Table ). Validated serologic tests for Bb exposure in North America include in‐house and reference laboratory C 6 ‐based testing (SNAP4Dx and SNAP4DxPlus [IDEXX Laboratories, Westbrook, Maine]; Lyme Quant C6 [IDEXX Laboratories, Westbrook, Maine]), the VetScan Canine Lyme Rapid assay (Abaxis North America, Union City, California), the AccuPlex4 test (Antech Diagnostics, Irvine, California), and the Multiplex test (Cornell University Animal Health Diagnostic Center, Ithaca, New York) . Few studies compare different Bb antibody assay performance.…”

“…Six‐month boosters have been proposed during the initial year (although no safety studies are available) and it is unknown whether or not to suggest 6‐month versus annual boosters thereafter. Vaccine failures were found only 22 weeks after OspA subunit or bacterin vaccination . The most recently licensed Bb vaccine (Vanguard crLyme, Zoetis, Florham Park, New Jersey 07932) was shown in prerelease studies to induce OspA and OspC antibodies against 7 Bb strains, which may afford broader protection.…”

“…[83][84][85]87 The OspC antibodies probably increase again in field conditions (ie, re-exposure is a natural booster), thus finding OspC antibodies in a nonvaccinated dog may indicate recent exposure or re-exposure, without specifying when the dog was first infected in its life. The OspA antibodies usually are a marker for vaccination, but they may develop transiently in early infection, 83,85 or possibly later during chronic infections, as seen in infected humans, because Bb displays antigenic variation, and expresses its antigenic repertoire over time to avoid host immunity. [88][89][90][91] The C 6 result has been shown to wane after treatment; [92][93][94] OspF antibodies also may wane.…”