Comparison of serological and molecular panels for diagnosis of vector\borne diseases in dogs. 1, San Antonio Type 2California 1), 3 subsp. genotypes (I, II, and III), ((spp. PCR\positive dogs were seroreactive to any of the 8 IFA antigens, indicating low IFA sensitivity. PCR\positive dogs were most often IFA seroreactive to (=?15), to II (=?13), or to both (= 9) antigens. Of the 26 previously IFA\unfavorable/PCR\unfavorable dogs, 4 (15%) were seroreactive using the expanded antigen panel. Conclusion and Clinical Importance Despite IFA testing of dogs against 8 different isolates, IFA sensitivity remained poor, and specificity was only 85%. Development of a reliable serological assay is needed to facilitate the diagnosis of contamination in dogs. alpha\proteobacteria growth mediumCAL1 California 1H1 Houston 1SA2 San Antonio 2Bh subspecies spp. are transmitted to mammals by arthropod vectors, including ticks, fleas, keds, lice, mites, and sand flies.1, 2, 3 Various arthropods transmit different spp. among reservoir and incidental hosts, thereby complicating and confounding clinical, diagnostic, and epidemiological analyses.4, 5, 6, 7 Currently, serology, as well as culture\based and polymerase chain reaction (PCR) assays, are relatively insensitive for PF-06424439 the diagnosis of bartonellosis in dogs.8, 9, 10 Immunofluorescent antibody assays (IFAs) are the most frequently used serological testing modalities for the diagnosis of bartonellosis in dogs.8, 11, 12 Studies PF-06424439 involving dogs, humans, and other animals have reported inconsistent and variable sensitivities and specificities for IFAs.13, 14, 15, 16, 17, 18 Genetically different spp. and strains are widespread in humans and animals throughout the world.19, 20, 21 Therefore, a possible explanation for variation among studies is exposure to spp., subspecies, or strain that differs from the IFA antigen used for diagnostic testing.12, 13, 16 Diagnostically important differences in serological responses have been documented in animals and human patients depending on Rabbit Polyclonal to CLNS1A which isolate/strain was used as an antigen.13, 16, 22 Further complicating diagnoses, clinical signs, pathologic sequelae, and antibody kinetics PF-06424439 can vary among individual animals infected with the same strain.23, 24, 25 Because members of the genus can induce long\lasting bacteremia, the stage of contamination (acute, subacute, or chronic) also contributes to variation in antibody detection.23, 24 Subjectivity associated with IFA interpretation and variability in technical variables among laboratories further contribute to differences in antibody detection or reported antibody titers. Thus, spp. serodiagnosis is usually influenced by variations in bacterial, host, and laboratory variables. Although 10 spp. have been implicated in association with endocarditis, myocarditis, or other disease manifestations in dogs; ((subsp. (represented the first spp. isolated from dogs.26 Therefore, initial IFA testing used as the sole antigen source for diagnostic and research purposes. Subsequently, 4 genotypes and several other spp. were found to infect dogs, including and spp., dogs develop a species\specific IFA antibody response.12, 28 However, bacteremic sick dogs frequently are seroreactive to multiple IFA antigens or alternatively they are not spp. seroreactive despite extended illness durations.8, 15 Ideally, a serological assay used for PF-06424439 epidemiological or diagnostic purposes should detect antibodies regardless of the infecting spp., genotype, or strain. Currently, because there are at least 38 named and Candidatus spp., with nearly half implicated in association with infections of dogs or humans, we posed the question: Would a broader panel of spp. antigens increase the serodiagnostic sensitivity and specificity of IFAs? We hypothesized that a comprehensive panel of spp. isolates would increase IFA serodiagnostic sensitivity, while optimizing specificity. Therefore, the purpose of our study was to evaluate the sensitivity and specificity of 8 IFAs using archived serum samples from spp. naturally\uncovered (PCR\positive) and presumptively non\uncovered (seronegative/PCR\unfavorable) dogs. 2.?MATERIALS AND METHODS 2.1. Source of sera for immunofluorescent antibody assays testing Sixty archived sera from PF-06424439 dogs previously tested at the North Carolina State University, College of Veterinary Medicine, Vector Borne Diseases Diagnostic Laboratory (NCSU\CVM\VBDDL) were selected for comparative IFA testing against 8 cell culture\produced spp. antigens. Serum samples were categorized into 2 groups to assess sensitivity and specificity. All sera were submitted to the NCSU\CVM\VBDDL for diagnostic testing between 2011 and 2016. After initial processing by the NCSU\CVM\VBDDL, sera were stored at ?80?C. 2.1.1. Group I (polymerase chain reaction positive dogs) Group I consisted of 34 stored frozen serum samples from spp. naturally infected dogs (PCR\positive) for which the species, genotype, or strain was confirmed by DNA sequencing. We could only identify.
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