RT info:eu-repo/semantics/article T1 A molecular toolbox for fast and convenient diagnosis of emerging and reemerging bacterial pathogens causing fever of intermediate duration. A1 Carmelo, Emma A1 Vega García, Elva del Cristo A1 Palacios, Génesis A1 Pérez, José A. A1 Vélez‑Tobarias, Mónica A1 Torres‑Vega, Ana María A1 Ascaso‑Terrén, Carlos A2 Obstetricia y GinecologíaPediatría, Medicina Preventiva y Salud Pública, Toxicología y Medicina Legal y Forense y Parasitología A2 Investigación en Parasitología K1 Molecular toolbox  K1 Fever of intermediate duration K1 Arthropod-borne bacteria K1 Real-time PCR K1 FRET probes K1 Molecular diagnostics AB Purpose Fever of intermediate duration (FID) is defined as a fever in the community without a specific origin or focus, with a duration between 7 and 28 days. 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 Rickettsia, Bartonella, Anaplasma, and Ehrlichia, as well as Coxiella burnetii. Methods Reference DNA sequences from a collection of Rickettsia, Bartonella, Anaplasma, and Ehrlichia species were used to design genus-specific primers and FRET probes targeted to conserved genomic regions. For C. burnetii, 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. Results The four real-time PCR assays can detect as few as ten copies of target DNA from those five genera of FDIcausing bacteria in a background of 300 ng of human genomic DNA, mimicking the low microbial load generally found in patient’s blood. Conclusion These assays constitute a fast and convenient “toolbox” 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. YR 2024 FD 2024 LK http://riull.ull.es/xmlui/handle/915/37290 UL http://riull.ull.es/xmlui/handle/915/37290 LA en DS Repositorio institucional de la Universidad de La Laguna RD 05-nov-2024