{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T08:13:36Z","timestamp":1790669616677,"version":"4.1.0"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2017,9,1]],"date-time":"2017-09-01T00:00:00Z","timestamp":1504224000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2017,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The emergence of fluoroquinolone (FQ)-resistant mutants of\n            <jats:named-content content-type=\"genus-species\">Legionella pneumophila<\/jats:named-content>\n            in infected humans was previously reported using a next-generation DNA sequencing (NGS) approach. This finding could explain part of the therapeutic failures observed in legionellosis patients treated with these antibiotics. The aim of this study was to develop digital PCR (dPCR) assays allowing rapid and accurate detection and quantification of these resistant mutants in respiratory samples, especially when the proportion of mutants in a wild-type background is low. We designed three dPCRgyrA assays to detect and differentiate the wild-type and one of the three\n            <jats:italic>gyrA<\/jats:italic>\n            mutations previously described as associated with FQ resistance in\n            <jats:named-content content-type=\"genus-species\">L. pneumophila<\/jats:named-content>\n            : at positions 248C\u2192T (T83I), 259G\u2192A (D87N), and 259G\u2192C (D87H). To assess the performance of these assays, mixtures of FQ-resistant and -susceptible strains of\n            <jats:named-content content-type=\"genus-species\">L. pneumophila<\/jats:named-content>\n            were analyzed, and the results were compared with those obtained with Sanger DNA sequencing and real-time quantitative PCR (qPCR) technologies. The dPCRgyrA assays were able to detect mutated\n            <jats:italic>gyrA<\/jats:italic>\n            sequences in the presence of wild-type sequences at up to 1:1,000 resistant\/susceptible allele ratios. By comparison, Sanger DNA sequencing and qPCR were less sensitive, allowing the detection of\n            <jats:italic>gyrA<\/jats:italic>\n            mutants at up to 1:1 and 1:10 ratios, respectively. When testing 38 respiratory samples from 23 legionellosis patients (69.6% treated with an FQ), dPCRgyrA detected small amounts of\n            <jats:italic>gyrA<\/jats:italic>\n            mutants in four (10.5%) samples from three (13.0%) patients. These results demonstrate that dPCR is a highly sensitive alternative to quantify FQ resistance in\n            <jats:named-content content-type=\"genus-species\">L. pneumophila<\/jats:named-content>\n            , and it could be used in clinical practice to detect patients that could be at higher risk of therapeutic failure.\n          <\/jats:p>","DOI":"10.1128\/aac.00628-17","type":"journal-article","created":{"date-parts":[[2017,7,4]],"date-time":"2017-07-04T00:40:52Z","timestamp":1499128852000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":23,"title":["Digital PCR for Detection and Quantification of Fluoroquinolone Resistance in Legionella pneumophila"],"prefix":"10.1128","volume":"61","author":[{"given":"Aur\u00e9lie","family":"Hennebique","sequence":"first","affiliation":[{"name":"Institut de Biologie et de Pathologie, Centre Hospitalier Universitaire Grenoble Alpes, Grenoble, France"},{"name":"Universit\u00e9 Grenoble Alpes, TheREx, TIMC-IMAG, CNRS, Grenoble, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Bidart","sequence":"additional","affiliation":[{"name":"Institut de Biologie et de Pathologie, Centre Hospitalier Universitaire Grenoble Alpes, Grenoble, France"},{"name":"Universit\u00e9 Grenoble Alpes, Braintech Lab, CEA, INSERM, Grenoble, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Jarraud","sequence":"additional","affiliation":[{"name":"Centre National de R\u00e9f\u00e9rence des L\u00e9gionelles, Institut des Agents Infectieux, Groupe Hospitalier Nord, Hospices Civils de Lyon, Lyon, France"},{"name":"Centre International de Recherche en Infectiologie, INSERM, Universit\u00e9 Lyon 1, CNRS, ENS, Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"La\u00ebtitia","family":"Beraud","sequence":"additional","affiliation":[{"name":"Centre National de R\u00e9f\u00e9rence des L\u00e9gionelles, Institut des Agents Infectieux, Groupe Hospitalier Nord, Hospices Civils de Lyon, Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carole","family":"Schwebel","sequence":"additional","affiliation":[{"name":"R\u00e9animation, Centre Hospitalier Universitaire Grenoble Alpes, Grenoble, France"},{"name":"Radiopharmaceutiques Biocliniques, INSERM, La Tronche, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Max","family":"Maurin","sequence":"additional","affiliation":[{"name":"Institut de Biologie et de Pathologie, Centre Hospitalier Universitaire Grenoble Alpes, Grenoble, France"},{"name":"Universit\u00e9 Grenoble Alpes, TheREx, TIMC-IMAG, CNRS, Grenoble, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandrine","family":"Boisset","sequence":"additional","affiliation":[{"name":"Institut de Biologie et de Pathologie, Centre Hospitalier Universitaire Grenoble Alpes, Grenoble, France"},{"name":"Universit\u00e9 Grenoble Alpes, TheREx, TIMC-IMAG, CNRS, Grenoble, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1086\/428059"},{"key":"e_1_3_3_3_2","first-page":"495","article-title":"Macrolides versus quinolones in Legionella pneumonia: results from the Community-Acquired Pneumonia Organization international study","volume":"14","author":"Griffin AT","year":"2010","unstructured":"Griffin AT PeyraniP WiemkenT ArnoldF. 2010. Macrolides versus quinolones in Legionella pneumonia: results from the Community-Acquired Pneumonia Organization international study. Int J Tuberc Lung Dis14:495\u2013499.","journal-title":"Int J Tuberc Lung Dis"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/22.3.389"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.11.4294-4295.2000"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1086\/429826"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0732-8893(99)00095-4"},{"key":"e_1_3_3_8_2","first-page":"137","article-title":"The amino acid substitution Asp87Tyr in GyrA of Legionella pneumophila Sg 1 is associated with elevated MICs to nalidixic acid but not to ciprofloxacin or moxifloxacin in vitro","volume":"292","author":"Linde H","year":"2002","unstructured":"Linde H MorgenrothK BarabasS LehnN. 2002. The amino acid substitution Asp87Tyr in GyrA of Legionella pneumophila Sg 1 is associated with elevated MICs to nalidixic acid but not to ciprofloxacin or moxifloxacin in vitro. 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