{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:54:44Z","timestamp":1785430484728,"version":"3.56.0"},"reference-count":36,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T00:00:00Z","timestamp":1765929600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T00:00:00Z","timestamp":1765929600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100001458","name":"American Kennel Club Canine Health Foundation","doi-asserted-by":"publisher","award":["02287-E"],"award-info":[{"award-number":["02287-E"]}],"id":[{"id":"10.13039\/100001458","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100011490","name":"College of Veterinary Medicine, North Carolina State University","doi-asserted-by":"publisher","award":["Bartonella\/Vector Borne Disease Research Fund"],"award-info":[{"award-number":["Bartonella\/Vector Borne Disease Research Fund"]}],"id":[{"id":"10.13039\/100011490","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2025,12,17]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                      species, emerging vector-borne pathogens of dogs, are increasingly associated with severe, chronic sequelae, as well as potentially life-threatening diseases, such as endocarditis and myocarditis. Diagnosis of bartonelloses is mainly based on PCR, culture, and serological assays. Despite molecular and biotechnological advances, serological assays employing immunofluorescence antibody (IFA), Western blotting, and enzyme-linked immunosorbent assay (ELISA) technologies have encountered diagnostic limitations, primarily due to poor sensitivity. Using sera from\n                      <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                      -infected and na\u00efve dogs, we applied an immunoproteomic approach to develop a reliable ELISA assay for the diagnosis of bartonelloses in dogs. Five recombinant\n                      <jats:italic toggle=\"yes\">Bartonella henselae<\/jats:italic>\n                      immunodominant proteins (rATP-\u03b2, rGroEL, rLemA, rSucB, and rVirB5) were tested in an ELISA format. Sensitivity and specificity of each protein were calculated based on an imperfect reference IFA assay. Dogs comprised Group I: 36\n                      <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                      spp. naturally infected dogs (all\n                      <jats:italic toggle=\"yes\">B. henselae<\/jats:italic>\n                      IFA seroreactive) and Group II: 34\n                      <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                      spp. PCR-negative and IFA-negative dogs. Based upon the ELISA seroreactivity results, rATP-\u03b2 and rGroEL represented the most sensitive and specific candidate peptide targets for utilization in a canine diagnostic ELISA assay. rGroEL resulted in the sensitivity of 83% and specificity of 94% at an optical density (OD) cutoff value of 0.439 and area under curve (AUC) score of 0.93 (95% CI 0.87\u20130.99), while the sensitivity and specificity of rATP-\u03b2 were 69% and 94%, respectively, at a cutoff value of 0.565. The combination of rATP-\u03b2 with rGroEL resulted in an improved sensitivity of 88% and specificity of 92% at an OD cutoff value of 0.505. A receiver operating characteristic curve analysis for the rATP-\u03b2 plus rGroEL yielded an AUC score of 0.899 (95% CI 0.809\u20130.989). Combining rATP-\u03b2 with rGroEL could potentially further improve both the diagnostic sensitivity and specificity of an ELISA assay for the diagnosis of canine bartonelloses.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                        species are associated with a wide spectrum of clinical signs and life-threatening diseases in dogs. There is an increased risk of\n                        <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                        transmission from dogs to dogs, and from dogs to other animals and humans via vectors, such as ticks, fleas, or direct contact with infected clinical specimens. Due to the poor sensitivity of currently available molecular and serological assays, the diagnosis, treatment, and prevention of\n                        <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                        infection in dogs remains challenging. Developing a reliable serodiagnostic assay is essential for the clinical management of canine bartonelloses, a group of infections caused by\n                        <jats:italic toggle=\"yes\">Bartonella<\/jats:italic>\n                        species in dogs. Rapid diagnosis and timely treatment of canine bartonelloses could save the lives of thousands of dogs worldwide each year. This study provides key insights into the design of diagnostic tools utilizing\n                        <jats:italic toggle=\"yes\">Bartonella henselae<\/jats:italic>\n                        proteins that show promise as serological markers to improve the diagnosis of canine bartonelloses.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jcm.00267-25","type":"journal-article","created":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T17:21:43Z","timestamp":1764177703000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Development and validation of enzyme-linked immunosorbent assays for the serodiagnosis of canine 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