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Article

Comparison of Serological and Molecular Assays for Bartonella Species in Dogs with Hemangiosarcoma

1
Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53713, USA
2
Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA
3
Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Lawrence S. Young
Pathogens 2021, 10(7), 794; https://doi.org/10.3390/pathogens10070794
Received: 21 May 2021 / Revised: 18 June 2021 / Accepted: 18 June 2021 / Published: 23 June 2021
(This article belongs to the Special Issue The Evolving Biomedical Importance of Bartonella Species Infections)
Currently, a gold standard diagnostic test for Bartonella infection in dogs is lacking. This represents a critical limitation for the development and evaluation of new diagnostic tests, as well as for the diagnosis of, and research on, bartonellosis in dogs. This retrospective observational study aims to compare the results of commonly performed and newly-reported Bartonella spp. diagnostic tests in banked clinical specimens from 90 dogs with hemangiosarcoma (HSA) using composite reference standard (CRS) and random effects latent class analysis (RE-LCA) techniques. Samples from each dog were tested using six serological or molecular diagnostic assays, including indirect fluorescent antibody (IFA) and Western blot (WB) for the detection of antibodies in serum, and qPCR and droplet digital PCR (ddPCR) in blood and fresh frozen tissue biopsy samples (mainly splenic HSA tumors and histopathologically normal spleen or skin/adipose tissue). Bartonella infection prevalence was estimated to be 78% based on the CRS (parallel testing with all six assays), and 64% based on the RE-LCA model. The assay with the highest diagnostic accuracy was qPCR performed on fresh frozen tissue biopsy samples (sensitivity: 94% by RE-LCA and 80% by CRS; specificity: 100%). When comparing newly-reported to traditional Bartonella diagnostic assays, ddPCR was more sensitive for the detection of Bartonella DNA than qPCR when testing blood samples (36% vs. 0%, p < 0.0001). Dogs that were positive on serological assays alone with negative molecular assays were highly unlikely (<3%) to be classified as infected by the RE-LCA model. These data indicate that Bartonella spp. DNA can be PCR amplified from fresh frozen tissues from a majority of dogs with HSA using both qPCR and ddPCR, supporting the use of these methods for future controlled studies comparing the prevalence of Bartonella spp. DNA in the tissue of dogs with HSA to that of unaffected controls. View Full-Text
Keywords: diagnostic testing; ddPCR; PCR; serology; sensitivity; specificity; tissue biopsy; latent class analysis diagnostic testing; ddPCR; PCR; serology; sensitivity; specificity; tissue biopsy; latent class analysis
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MDPI and ACS Style

Lashnits, E.; Neupane, P.; Bradley, J.M.; Richardson, T.; Maggi, R.G.; Breitschwerdt, E.B. Comparison of Serological and Molecular Assays for Bartonella Species in Dogs with Hemangiosarcoma. Pathogens 2021, 10, 794. https://doi.org/10.3390/pathogens10070794

AMA Style

Lashnits E, Neupane P, Bradley JM, Richardson T, Maggi RG, Breitschwerdt EB. Comparison of Serological and Molecular Assays for Bartonella Species in Dogs with Hemangiosarcoma. Pathogens. 2021; 10(7):794. https://doi.org/10.3390/pathogens10070794

Chicago/Turabian Style

Lashnits, Erin, Pradeep Neupane, Julie M. Bradley, Toni Richardson, Ricardo G. Maggi, and Edward B. Breitschwerdt 2021. "Comparison of Serological and Molecular Assays for Bartonella Species in Dogs with Hemangiosarcoma" Pathogens 10, no. 7: 794. https://doi.org/10.3390/pathogens10070794

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