{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T20:33:02Z","timestamp":1779309182137,"version":"3.51.4"},"reference-count":34,"publisher":"Microbiology Society","issue":"7","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,7,23]]},"abstract":"\n Introduction.<\/jats:bold> Bartonellosis is an emerging zoonotic disease caused by bacteria of the genus \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>. Mixed \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> infections are a well-documented phenomenon in mammals and their ectoparasites. The accurate identification of \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> species in single and mixed infections is valuable, as different \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> species have varying impacts on infected hosts.<\/jats:p>\n \n Gap Statement.<\/jats:bold> Current diagnostic methods are inadequate at identifying the \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> species present in mixed infections.<\/jats:p>\n \n Aim.<\/jats:bold> The aim of this study was to adopt a Next Generation Sequencing (NGS) approach using Illumina sequencing technology to identify \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> species and demonstrate that this approach can resolve mixed \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> infections.<\/jats:p>\n \n Methodology.<\/jats:bold> We used Illumina PCR amplicon NGS to target the ssrA<\/jats:italic> and gltA<\/jats:italic> genes of \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> in fleas collected from cats, dogs and a hedgehog in Israel. We included artificially mixed \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> samples to demonstrate the ability for NGS to resolve mixed infections and we compared NGS to traditional Sanger sequencing.<\/jats:p>\n \n Results.<\/jats:bold> In total, we identified 74 Ctenocephalides felis<\/jats:italic>, two Ctenocephalides canis,<\/jats:italic> two Pulex irritans<\/jats:italic> and three Archaeopsylla e. erinacei<\/jats:italic> fleas. Real-time PCR of a subset of 48 fleas revealed that twelve were positive for \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>, all of which were cat fleas. Sanger sequencing of the ssrA<\/jats:italic> and gltA<\/jats:italic> genes confirmed the presence of \n \n Bartonella henselae<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>, \n \n Bartonella clarridgeiae<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> and \n \n Bartonella koehlerae<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>. Illumina NGS of ssrA<\/jats:italic> and gltA<\/jats:italic> amplicons further confirmed the \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> species identity in all 12 flea samples and unambiguously resolved the artificially mixed \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> samples.<\/jats:p>\n \n Conclusion.<\/jats:bold> The adaptation and multiplexing of existing PCR assays for diversity profiling via NGS is a feasible approach that is superior to traditional Sanger sequencing for \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> speciation and resolving mixed \n \n Bartonella<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic> infections. The adaptation of other PCR primers for Illumina NGS will be useful in future studies where mixed bacterial infections may be present.<\/jats:p>","DOI":"10.1099\/jmm.0.001400","type":"journal-article","created":{"date-parts":[[2021,7,23]],"date-time":"2021-07-23T13:15:54Z","timestamp":1627046154000},"source":"Crossref","is-referenced-by-count":6,"title":["Adaptation of gltA and ssrA assays for diversity profiling by Illumina sequencing to identify Bartonella henselae, B. clarridgeiae and B. koehlerae"],"prefix":"10.1099","volume":"70","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3271-8925","authenticated-orcid":false,"given":"Rosemonde Isabella","family":"Power","sequence":"first","affiliation":[{"name":"Sydney School of Veterinary Science, Faculty of Science, University of Sydney, Sydney 2006, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8787-6755","authenticated-orcid":false,"given":"Nichola Elisa Davies","family":"Calvani","sequence":"additional","affiliation":[{"name":"Sydney School of Veterinary Science, Faculty of Science, University of Sydney, Sydney 2006, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7336-4698","authenticated-orcid":false,"given":"Yaarit","family":"Nachum-Biala","sequence":"additional","affiliation":[{"name":"Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, Rehovot 7610001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3250-883X","authenticated-orcid":false,"given":"Harold","family":"Salant","sequence":"additional","affiliation":[{"name":"Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, Rehovot 7610001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0542-207X","authenticated-orcid":false,"given":"Shimon","family":"Harrus","sequence":"additional","affiliation":[{"name":"Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, Rehovot 7610001, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1459-9117","authenticated-orcid":false,"given":"Jan","family":"\u0160lapeta","sequence":"additional","affiliation":[{"name":"Sydney School of Veterinary Science, Faculty of Science, University of Sydney, Sydney 2006, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"publisher","DOI":"10.1186\/1297-9716-43-15"},{"key":"R2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00013-17"},{"key":"R3","doi-asserted-by":"publisher","DOI":"10.1001\/jama.271.7.531"},{"key":"R4","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1086\/514671","article-title":"Chest-wall abscess due to cat-scratch disease (CSD) in an adult with antibodies to Bartonella clarridgeiae: Case report and review of the thoracopulmonary manifestations of CSD","volume":"27","author":"Margileth","year":"1998","journal-title":"Clin Infect Dis"},{"key":"R5","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.7.1813-1818.1997"},{"key":"R6","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.42.8.3462-3468.2004"},{"key":"R7","doi-asserted-by":"publisher","DOI":"10.1093\/jmedent\/33.3.490"},{"key":"R8","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.3.1876-1879.2001"},{"key":"R9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0004128"},{"key":"R10","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.34.8.1952-1956.1996"},{"key":"R11","doi-asserted-by":"publisher","DOI":"10.1186\/s13071-017-2372-5"},{"key":"R12","first-page":"36","article-title":"Molecular identity of cat fleas (Ctenocephalides felis) from cats in Georgia, USA carrying Bartonella clarridgeiae, Bartonella henselae and Rickettsia sp. 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