Comparison of indirect immunofluorescent-antibody assay, enzyme-linked immunosorbent assay, and Western immunoblot for the diagnosis of Lyme disease in dogs

Abstract

Enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescent-antibody assay (IFA), and Western immunoblot were used to test serum samples from 128 dogs for the presence of antibody to Borrelia burgdorferi. Sera included 72 samples from dogs suspected of having Lyme disease, 32 samples from dogs residing in areas in which Lyme disease was not considered endemic, and 24 samples from dogs with clinical and serologic evidence of immune-mediated disease (n = 10), Rocky Mountain spotted fever (n = 5), or leptospirosis (n = 9). Results of Western immunoblotting were used as the standard against which performances of ELISA and IFA were measured. ELISA was significantly more sensitive than IFA (84.8 versus 66.7%), although both tests were equally specific (93.5%). Eight samples that were positive by Western immunoblot were simultaneously negative by ELISA and IFA. Of these eight, four were from dogs suspected of having immune-mediated disease, two were from dogs suspected of having leptospirosis, and two were from dogs suspected of having Lyme disease. These results may indicate that sera from dogs with immune-mediated disease, and to a lesser extent sera from those with leptospirosis, cross-react with B. burgdorferi antigens. Alternatively, Western immunoblot results may not truly reflect Lyme disease status, particularly in the case of dogs with immune-mediated diseases. At present, however, the use of Western immunoblotting as a diagnostic standard for dogs offers the best alternative to a clinical definition of disease.

Cross-reactivity between B. burgdorferi and other spirochetes affects specificity of serotests for detection of antibodies to the Lyme disease agent in dogs

“…Western immunoblotting has been used to analyze the antibody response or to confirm Lyme disease infection in human patients (Karlsson et al, 1989;Karlsson, 1990;Zoller et al, 1991 ). Western blotting has confirmed KELA results in our laboratory and has also been used elsewhere as a confirmatory test for dog Lyme disease Lindenmayer et al, 1990;Jacobson, et al, 1992).…”

“…these include the indirect immunofluorescence assay (IFA), kinetics-based enzyme-linked immunosorbent assay (KELA), and Western blot (Lindenmayer et al, 1990;Karlsson, 1990;Karlsson et al, 1989;Zoller et al, 1991 ). The specificity of serotest results for dogs, both in surveillance and clinical applications, thus becomes an important issue.…”

Borrelia burgdorferiInfection and Lyme Disease in North American Horses: A Consensus Statement

“…29,63 Whole cell lysate assays such as IFAT and ELISA do not distinguish between infection stages or between natural infection and vaccination. Additionally, evidence from dogs and horses suggests that whole cell lysate assays can give false positive results attributable to crossreactivity with antibodies against common bacterial antigens such as flagellar proteins, [67][68][69] and positive IFAT or ELISA results therefore require confirmation of infection by WB. Serologic surveys have demonstrated that clinically normal horses living in endemic areas often have detectable antibody levels against B. burgdorferi.…”

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

“…Since Borrelia burgdorferi was found to be the causative agent of Lyme disease, various methods have been employed for the determination of antibodies to the spirochete in humans and in domestic and wild animals. The enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent-antibody assay (IFA) have been used to screen serum, and immunoblotting techniques have been used to confirm positive results (1,5,10,15,21,22,31). Various studies have determined the type and number of bands that must be present for a sample to be considered positive (5,17,24,33) and to distinguish between the early and late stages of Lyme disease in humans (32).…”