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FID is often caused by pathogens associated with animal contact or their arthropods parasites, such as ticks, fleas, or lice. The purpose of this work is to design a collection of molecular tools to promptly and accurately detect common bacterial pathogens causing FID, including bacteria belonging to genera <jats:italic>Rickettsia<\/jats:italic>, <jats:italic>Bartonella<\/jats:italic>, <jats:italic>Anaplasma<\/jats:italic>, and <jats:italic>Ehrlichia<\/jats:italic>, as well as<jats:italic> Coxiella burnetii.<\/jats:italic><\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>Reference DNA sequences from a collection of <jats:italic>Rickettsia<\/jats:italic>, <jats:italic>Bartonella<\/jats:italic>, <jats:italic>Anaplasma<\/jats:italic>, and <jats:italic>Ehrlichia<\/jats:italic> species were used to design genus-specific primers and FRET probes targeted to conserved genomic regions. For <jats:italic>C. burnetii<\/jats:italic>, primers previously described were used, in combination with a newly designed specific probe. Real-time PCR assays were optimized using reference bacterial genomic DNA in a background of human genomic DNA.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>The four real-time PCR assays can detect as few as ten copies of target DNA from those five genera of FDI-causing bacteria in a background of 300\u00a0ng of human genomic DNA, mimicking the low microbial load generally found in patient\u2019s blood.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusion<\/jats:title>\n                <jats:p>These assays constitute a fast and convenient \u201ctoolbox\u201d that can be easily implemented in diagnostic laboratories to provide timely and accurate detection of bacterial pathogens that are typical etiological causes of febrile syndromes such as FID in humans.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1007\/s10096-024-04760-8","type":"journal-article","created":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T14:23:09Z","timestamp":1706192589000},"page":"649-657","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["A molecular toolbox for fast and convenient diagnosis of emerging and reemerging bacterial pathogens causing fever of intermediate duration"],"prefix":"10.1007","volume":"43","author":[{"given":"Elva","family":"Vega-Garc\u00eda","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e9nesis","family":"Palacios","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 A.","family":"P\u00e9rez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00f3nica","family":"V\u00e9lez-Tobarias","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Mar\u00eda","family":"Torres-Vega","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos","family":"Ascaso-Terr\u00e9n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1126-993X","authenticated-orcid":false,"given":"Emma","family":"Carmelo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,1,25]]},"reference":[{"key":"4760_CR1","doi-asserted-by":"publisher","first-page":"2047","DOI":"10.1007\/s10096-021-04322-2","volume":"40","author":"E Guirao-Arrabal","year":"2021","unstructured":"Guirao-Arrabal E, Mu\u00f1oz-Medina L, Anguita-Santos F, Vinuesa-Garc\u00eda D, Hern\u00e1ndez-Quero J (2021) Empirical treatment with doxycycline of fever of intermediate duration. 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