Validation of Bartonella henselae Western Immunoblotting for Serodiagnosis of Bartonelloses in Dogs

Pradeep Neupane; Sindhura Sevala; Nandhakumar Balakrishnan; Henry Marr; James Wilson; Ricardo Maggi; Adam Birkenheuer; Michael Lappin; Bruno Chomel; Edward B Breitschwerdt

doi:10.1128/JCM.01335-19

Validation of Bartonella henselae Western Immunoblotting for Serodiagnosis of Bartonelloses in Dogs

J Clin Microbiol. 2020 Mar 25;58(4):e01335-19. doi: 10.1128/JCM.01335-19. Print 2020 Mar 25.

Authors

Pradeep Neupane^{1

2}, Sindhura Sevala^{1

2}, Nandhakumar Balakrishnan³, Henry Marr^{1

2}, James Wilson^{1

2}, Ricardo Maggi^{1

2}, Adam Birkenheuer^{1

2}, Michael Lappin⁴, Bruno Chomel⁵, Edward B Breitschwerdt^{6

2}

Affiliations

¹ Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.
² Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.
³ Bureau of Laboratories, Division of Infectious Disease, Michigan Department of Health and Human Services, Lansing, Michigan, USA.
⁴ Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
⁵ Department of Population Health & Reproduction, School of Veterinary Medicine, University of California, Davis, California, USA.
⁶ Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA ebbreits@ncsu.edu.

Abstract

Bartonella spp. are etiological agents of life-threatening zoonotic diseases in dogs worldwide. Due to the poor sensitivity of immunofluorescent-antibody assays (IFAs), a reliable serodiagnostic test for canine bartonelloses is of clinical importance. The utility of Western blotting (WB) for the serodiagnosis of canine bartonelloses has not been critically investigated. The objective of this study was to characterize WB immunodominant proteins that could be used to confirm a serodiagnosis of bartonelloses. Using agar-grown Bartonella henselae San Antonio type 2 (SA2) whole-cell proteins, sera derived from four dog groups were tested by WB to assess immunodominant protein recognition patterns: group I consisted of 92 serum samples (10 preexposure and 82 postexposure serum samples) from 10 adult beagles experimentally inoculated with Bartonella spp., group II consisted of 36 serum samples from Bartonella PCR-positive naturally infected dogs, group III consisted of 26 serum samples from Bartonella PCR-negative and IFA-negative dogs, and group IV consisted of serum samples from 8 Brucella canis IFA-positive and 10 Rickettsia rickettsii IFA-positive dogs. Following experimental inoculation, 9 (90%) group I dogs were variably seroreactive to one or more of six specific immunodominant proteins (13, 17, 29, 50, 56, and 150 kDa). There was a strong but variable recognition of these proteins among 81% of group II dogs. In contrast, 24/26 group III dogs were not reactive to any immunodominant protein. In this study, the sensitivity and diagnostic accuracy of B. henselae SA2 WB were higher than those of B. henselae SA2 IFA testing. Some B. henselae SA2 immunodominant proteins were recognized by dogs experimentally and naturally infected with Bartonella spp. other than B. henselae Additional research is necessary to more fully define the utility of WB for the serodiagnosis of canine bartonelloses.

Keywords: Bartonella henselae; Western blotting; antigen; serology; vector borne; zoonosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Bacterial
Bartonella Infections* / diagnosis
Bartonella Infections* / veterinary
Bartonella henselae*
Bartonella*
Blotting, Western
Dog Diseases* / diagnosis
Dogs
Serologic Tests

Substances

Antibodies, Bacterial