{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T16:21:47Z","timestamp":1788884507945,"version":"build-2803163510"},"reference-count":49,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2020,12,24]],"date-time":"2020-12-24T00:00:00Z","timestamp":1608768000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006533","name":"Ministerstvo Zem\u011bd\u011blstv\u00ed","doi-asserted-by":"publisher","award":["MZE-RO0518"],"award-info":[{"award-number":["MZE-RO0518"]}],"id":[{"id":"10.13039\/501100006533","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009532","name":"Ministerstvo Vnitra \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["VI 20152020044"],"award-info":[{"award-number":["VI 20152020044"]}],"id":[{"id":"10.13039\/100009532","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009532","name":"Ministerstvo Vnitra \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["VH20172020012"],"award-info":[{"award-number":["VH20172020012"]}],"id":[{"id":"10.13039\/100009532","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014911","name":"Ministerstvo Obrany \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["VF20142015039"],"award-info":[{"award-number":["VF20142015039"]}],"id":[{"id":"10.13039\/501100014911","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Early detection of biohazardous bacteria that can be misused as biological weapons is one of the most important measures to prevent the spread and outbreak of biological warfare. For this reason, many instrument platforms need to be introduced into operation in the field of biological warfare detection. Therefore the purpose of this study is to establish a new detection panel for biothreat bacteria (Bacillus anthracis, Yersinia pestis, Francisella tularensis, and Brucella spp.) and confirm it by collaborative validation by using a multiplex oligonucleotide ligation followed by polymerase chain reaction and hybridization to microspheres by MagPix detection platform (MOL-PCR). Appropriate specific sequences in bacterial DNA were selected and tested to assemble the detection panel, and MOLigo probes (short specific oligonucleotides) were designed to show no cross-reactivity when tested between bacteria and to decrease the background signal measurement on the MagPix platform. During testing, sensitivity was assessed for all target bacteria using serially diluted DNA and was determined to be at least 0.5 ng\/\u00b5L. For use as a diagnostic kit and easier handling, the storage stability of ligation premixes (MOLigo probe mixes) was tested. This highly multiplex method can be used for rapid screening to prevent outbreaks arising from the use of bacterial strains for bioterrorism, because time of analysis take under 4 h.<\/jats:p>","DOI":"10.3390\/microorganisms9010038","type":"journal-article","created":{"date-parts":[[2020,12,24]],"date-time":"2020-12-24T22:56:45Z","timestamp":1608850605000},"page":"38","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Development and Inter-Laboratory Validation of Diagnostics Panel for Detection of Biothreat Bacteria Based on MOL-PCR Assay"],"prefix":"10.3390","volume":"9","author":[{"given":"Pavlina","family":"Jelinkova","sequence":"first","affiliation":[{"name":"Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296\/70, 621 00 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0890-5811","authenticated-orcid":false,"given":"Jakub","family":"Hrdy","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296\/70, 621 00 Brno, Czech Republic"},{"name":"Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 753\/5, 625 00 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jirina","family":"Markova","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296\/70, 621 00 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiri","family":"Dresler","sequence":"additional","affiliation":[{"name":"Military Health Institute, Military Medical Agency, Tychonova 1, 160 01 Prague 6, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petr","family":"Pajer","sequence":"additional","affiliation":[{"name":"Military Health Institute, Military Medical Agency, Tychonova 1, 160 01 Prague 6, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oto","family":"Pavlis","sequence":"additional","affiliation":[{"name":"Military Health Institute, Military Medical Agency, Tychonova 1, 160 01 Prague 6, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavel","family":"Branich","sequence":"additional","affiliation":[{"name":"Military Veterinary Institute, Opavska 29, 748 01 Hlucin, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela","family":"Borilova","sequence":"additional","affiliation":[{"name":"Department of Meat Hygiene and Technology, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Palackeho tr. 1946\/1, 612 42 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5444-5019","authenticated-orcid":false,"given":"Marketa","family":"Reichelova","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296\/70, 621 00 Brno, Czech Republic"},{"name":"Collection of Animal Pathogenic Microorganisms, Department of Bacteriology, Veterinary Research Institute, Hudcova 296\/70, 621 00 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0826-1075","authenticated-orcid":false,"given":"Vladimir","family":"Babak","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 296\/70, 621 00 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7144-6323","authenticated-orcid":false,"given":"Nikol","family":"Reslova","sequence":"additional","affiliation":[{"name":"Department of Botany and Zoology, Faculty of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2836-1644","authenticated-orcid":false,"given":"Petr","family":"Kralik","sequence":"additional","affiliation":[{"name":"Department of Meat Hygiene and Technology, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Palackeho tr. 1946\/1, 612 42 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,24]]},"reference":[{"key":"ref_1","first-page":"2","article-title":"Bio threat s bacterial warfare agents","volume":"2","author":"Nishanth","year":"2011","journal-title":"J. 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