{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T12:32:42Z","timestamp":1753878762810,"version":"3.41.2"},"reference-count":31,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2016,12,28]],"date-time":"2016-12-28T00:00:00Z","timestamp":1482883200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004385","name":"Universiteit Gent","doi-asserted-by":"publisher","award":["GOA 01G00408"],"award-info":[{"award-number":["GOA 01G00408"]}],"id":[{"id":"10.13039\/501100004385","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Fonds Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Helicobacter"],"published-print":{"date-parts":[[2017,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p><jats:italic>Helicobacter suis<\/jats:italic> is a very fastidious microorganism associated with gastritis, gastric ulcers, and mucosa\u2010associated lymphoid tissue lymphoma in humans. <jats:italic>In vitro<\/jats:italic> isolation of this agent from human patients has so far been unsuccessful.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and methods<\/jats:title><jats:p>A probe\u2010based real\u2010time <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> (<jats:styled-content style=\"fixed-case\">RT<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content>) for the rapid detection of <jats:italic>H.\u00a0suis<\/jats:italic> in gastric biopsies was developed. Secondly, a mouse\u2010passage\u2010based protocol was optimized for isolation of low numbers of viable <jats:italic>H.\u00a0suis<\/jats:italic> bacteria. Mice were inoculated with different numbers of viable <jats:italic>H.\u00a0suis<\/jats:italic> (10<jats:sup>2<\/jats:sup>\u201010<jats:sup>8<\/jats:sup>) and kept for 4\u00a0weeks to allow multiplication of this pathogen.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The probe\u2010based real\u2010time <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> (<jats:styled-content style=\"fixed-case\">RT<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content>) exhibited a high degree of diagnostic specificity and analytical sensitivity, high linear correlations (<jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup> between 0.995 and 0.999), and high amplification efficiencies (&gt;90%) for <jats:italic>H.\u00a0suis<\/jats:italic>. No cross\u2010reactivity was detected with human, porcine, non\u2010human primate, and murine <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> nor with <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> from other bacteria including <jats:italic>Helicobacter<\/jats:italic> spp. and <jats:italic>Campylobacter<\/jats:italic> spp. <jats:italic>H.\u00a0suis<\/jats:italic> was successfully re\u2010isolated from the stomach of mice inoculated with at least 10<jats:sup>4<\/jats:sup> viable <jats:italic>H.\u00a0suis<\/jats:italic>, using a biphasic medium (<jats:styled-content style=\"fixed-case\">pH<\/jats:styled-content> 5), consisting of <jats:italic>Brucella<\/jats:italic> agar with <jats:italic>Brucella<\/jats:italic> broth on top, both supplemented with vitox supplement, Campylobacter\u2010selective supplement, amphotericin (5\u00a0\u03bcg\/mL), HCl (0.05%), fetal bovine serum (20%), and linezolid (5\u00a0\u03bcg\/mL). Linezolid was necessary to inhibit proliferation of contaminants, including lactobacilli.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The methods described above can be implemented for detection or isolation of <jats:italic>H.\u00a0suis<\/jats:italic> from human gastric biopsies.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hel.12369","type":"journal-article","created":{"date-parts":[[2016,12,28]],"date-time":"2016-12-28T11:18:00Z","timestamp":1482923880000},"source":"Crossref","is-referenced-by-count":14,"title":["A novel isolation protocol and probe\u2010based <scp>RT<\/scp>\u2010<scp>PCR<\/scp> for diagnosis of gastric infections with the zoonotic pathogen <i>Helicobacter suis<\/i>"],"prefix":"10.1111","volume":"22","author":[{"given":"Caroline","family":"Blaecher","sequence":"first","affiliation":[{"name":"Department of Pathology, Bacteriology and Avian Diseases Faculty of Veterinary Medicine Ghent University  Merelbeke 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