Quantitative Approach for the Serodiagnosis of Canine Lyme Disease by the Immunoblot Procedure

Abstract

Serum samples obtained from healthy, asymptomatic dogs in areas of Wisconsin and northern Illinois where Lyme disease is endemic or nonendemic were assayed for antibodies to Borrelia burgdorferi by enzyme-linked immunosorbent assay (ELISA), and positive results were confirmed by immunoblot assay. We found that 56.9% (562 of 1,077) of the samples were positive by ELISA and 82.0% (461 of 562) were positive by immunoblotting. A logistic regression model was developed to distinguish between nonvaccinated dogs naturally infected with B. burgdorferi from areas where the disease is endemic and dogs from areas where the disease is nonendemic that were vaccinated against Lyme disease. Of the 18 protein bands analyzed, 8 were found to be significantly different (P < 0.05) between the two groups. p93, p34, p31, and p28 occurred with increased frequency in vaccinated dogs, while p58, p37, p35, and p30 occurred more frequently in naturally infected dogs. The logistic regression equation obtained was used to determine the probability of natural infection among vaccinated dogs residing in areas where the disease is endemic. Of 125 samples, 87.2% had a very low probability of natural infection and only 2.4% were highly likely to be infected. Logistic regression is a useful method for distinguishing between vaccinated and naturally infected dogs and predicting the serological status of vaccinated dogs from areas where Lyme disease is endemic.

Seroprevalence, geographic distribution and risk factor analysis of Borrelia burgdorferi sensu lato in naturally exposed dogs of Iran

“…OspA band was found in a relatively high proportion of our Western blot positive dogs (73%). This is in contrast with results obtained by Greene and Straubinber (2006), who mentioned it should not be normally detected in naturally infected dogs, and Guerra et al (2000) or Leschnik et al (2010), who detected it at low levels in dogs with natural infection (17 and 2% early infection (EI) -28% late infection (LI) in USA and Austria, respectively). BmpA (39 kDa) was found in our study in 55% of positive dogs, while other authors reported an occurrence of 88% (Guerra et al, 2000) and 26% (EI) -33% (LI) (Leschnik et al, 2010).…”

“…This is in contrast with results obtained by Greene and Straubinber (2006), who mentioned it should not be normally detected in naturally infected dogs, and Guerra et al (2000) or Leschnik et al (2010), who detected it at low levels in dogs with natural infection (17 and 2% early infection (EI) -28% late infection (LI) in USA and Austria, respectively). BmpA (39 kDa) was found in our study in 55% of positive dogs, while other authors reported an occurrence of 88% (Guerra et al, 2000) and 26% (EI) -33% (LI) (Leschnik et al, 2010). p58 and p18 bands, which were found in 50 and 9% of our positive cases, respectively, while being detected in 84 and 64% of cases, respectively, by Guerra et al (2000), have not been detected at all by Leschnik et al (2010).…”

“…Reactivity, particularly to OspA (31 kDa) and OspB (34 kDa), possibly OspF (28 kDa), and 93 kDa bands occurs in vaccinated or parenterally inoculated dogs, but is normally absent or occurs (rarely) very late in naturally infected dogs (Greene and Straubinber, 2006). In our study, we found strongest antibody responses against flagellin, OspC and OspA (Figure 3), whereas Guerra et al (2000) and Leschnik et al (2010), who reported similar values for flagellin, showed quite lower values for OspC and OspA. The latter study in Austria was conducted using the same Western blot kit (MegaBlot) as we used.…”

“…Diagnosis of canine borreliosis can be made based on evidence of tick exposure in an endemic area, compatible clinical symptoms, detection of antibodies in blood serum, explicit response to antibiotics, and exclusion of other diseases (Fritz and Kjemtrup, 2003;Skotarczak, 2002;Speck et al, 2007). Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA) have been used as screening tests for detection of antibodies against B. burgdorferi sensu lato and immunoblotting procedure to confirm positive results (Guerra et al, 2000(Guerra et al, , 2001Lindenmayer et al, 1990).…”

ACVIM consensus update on Lyme borreliosis in dogs and cats

“…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. 95 Determination of quantitative titers to C 6 (or potentially OspF), pre-and 3 94 to 6 months post-treatment, were recommended by 4/6 panelists to check for a decrease after treatment as an indicator of decreased antigenic load, and to establish a new baseline for future comparison, because qualitative tests may stay positive a long time after treatment.…”