{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T12:47:04Z","timestamp":1787662024164,"version":"build-2736575974"},"reference-count":16,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,11,10]],"date-time":"2021-11-10T00:00:00Z","timestamp":1636502400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007356","name":"M\u00e9diterran\u00e9e Infection Foundation","doi-asserted-by":"publisher","award":["10-IAHU-03"],"award-info":[{"award-number":["10-IAHU-03"]}],"id":[{"id":"10.13039\/100007356","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Here, we develop a robust and sensitive real-time PCR assay which allows the simultaneous detection of vanA and vanB genes using common primers. The system was designed using the Primer3 online software. The specificity of primers and probes was first checked by in silico PCR and by BlastN analysis. The genomic DNA of 255 bacterial isolates, including Enterococcus spp., Gram-negative, and Gram-positive strains, as well as a collection of 50 stool and 50 rectal swab samples, were tested to evaluate the specificity of the new real-time PCR (RT-PCR) system. The results of the designed RT-PCR were 100% specific and 100% positive on tested vancomycin resistant isolates harboring either the vanA or vanB gene. RT-PCR assays were negative for all other bacterial species tested including vancomycin-sensitive Enterococci and Enterococcus strains harboring vanC genes. The limit of detection of vanA and vanB genes by RT-PCR assay was 47 CFU\/mL and 32 CFU\/mL, respectively. The rapid and accurate detection of vancomycin-resistant Enterococci is the cornerstone for minimizing the risk of nosocomial transmissions and outbreaks. We believe that this assay will strengthen routine diagnostics and surveillance programs.<\/jats:p>","DOI":"10.3390\/diagnostics11112081","type":"journal-article","created":{"date-parts":[[2021,11,10]],"date-time":"2021-11-10T09:19:21Z","timestamp":1636535961000},"page":"2081","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Real-Time PCR Assay for Rapid and Simultaneous Detection of vanA and vanB Genes in Clinical Strains"],"prefix":"10.3390","volume":"11","author":[{"given":"Hanane","family":"Zerrouki","sequence":"first","affiliation":[{"name":"Laboratoire de microbiologie appliqu\u00e9e \u00e0 l\u2019agroalimentaire, au biom\u00e9dical et \u00e0 l\u2019environnement, Universit\u00e9 de Tlemcen, Tlemcen 13000, Algeria"},{"name":"Aix-Marseille Univ., MEPHI, IRD, APHM, IHU-M\u00e9diterran\u00e9e Infection, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sid-Ahmed","family":"Rebiahi","sequence":"additional","affiliation":[{"name":"Laboratoire de microbiologie appliqu\u00e9e \u00e0 l\u2019agroalimentaire, au biom\u00e9dical et \u00e0 l\u2019environnement, Universit\u00e9 de Tlemcen, Tlemcen 13000, Algeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linda","family":"Hadjadj","sequence":"additional","affiliation":[{"name":"Aix-Marseille Univ., MEPHI, IRD, APHM, IHU-M\u00e9diterran\u00e9e Infection, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Marc","family":"Rolain","sequence":"additional","affiliation":[{"name":"Aix-Marseille Univ., MEPHI, IRD, APHM, IHU-M\u00e9diterran\u00e9e Infection, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6466-2324","authenticated-orcid":false,"given":"Seydina M.","family":"Diene","sequence":"additional","affiliation":[{"name":"Aix-Marseille Univ., MEPHI, IRD, APHM, IHU-M\u00e9diterran\u00e9e Infection, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2436","DOI":"10.1128\/JCM.00211-16","article-title":"Resistance mechanisms, epidemiology, and approaches to screening for vancomycin-resistant Enterococcus in the health care setting","volume":"54","author":"Faron","year":"2016","journal-title":"J. Clin. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.jhin.2014.09.002","article-title":"Controlling the spread of vancomycin-resistant enterococci. Is active screening worthwhile?","volume":"88","author":"Humphreys","year":"2014","journal-title":"J. Hosp. Infect."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1016\/j.idc.2016.07.009","article-title":"Vancomycin-Resistant Enterococci: Epidemiology, Infection Prevention, and Control","volume":"30","author":"Reyes","year":"2016","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1016\/j.mib.2010.08.004","article-title":"Horizontal gene transfer and the genomics of enterococcal antibiotic resistance","volume":"13","author":"Palmer","year":"2010","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_5","first-page":"788","article-title":"Global emergence and dissemination of enterococci as nosocomial pathogens: Attack of the clones?","volume":"7","author":"Rogers","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3351","DOI":"10.1093\/jac\/dkw312","article-title":"Multilevel population genetic analysis of vanA and vanB Enterococcus faecium causing nosocomial outbreaks in 27 countries (1986\u20132012)","volume":"71","author":"Freitas","year":"2016","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.idh.2018.12.002","article-title":"An outbreak of vanA vancomycin-resistant Enterococcus faecium in a hospital with endemic vanB VRE","volume":"24","author":"Hughes","year":"2019","journal-title":"Infect. Dis. Health"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Rangberg, A., Larsen, A.L., Kacelnik, O., S\u00e6ther, H.S., Bj\u00f8rland, M., Ringstad, J., and Jonassen, C.M. (2019). Molecular analysis and epidemiological typing of Vancomycin-resistant Enterococcus outbreak strains. Sci. Rep.","DOI":"10.1038\/s41598-019-48436-2"},{"key":"ref_9","first-page":"11917","article-title":"Real-time PCR for the rapid detection of vanA, vanB and vanM genes","volume":"9","author":"He","year":"2019","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1280","DOI":"10.1128\/jcm.31.5.1280-1285.1993","article-title":"Epidemiologic analysis and genotypic characterization of a nosocomial outbreak of vancomycin-resistant enterococci","volume":"31","author":"Boyle","year":"1993","journal-title":"J. Clin. 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Infect."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2636","DOI":"10.1128\/JCM.42.6.2636-2643.2004","article-title":"Comparison of the Roche LightCycler vanA\/vanB detection assay and culture for detection of vancomycin-resistant enterococci from perianal swabs","volume":"42","author":"Sloan","year":"2004","journal-title":"J. Clin. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1093\/jac\/dki029","article-title":"Characterization of a Tn5382-like transposon containing the vanB2 gene cluster in a Clostridium strain isolated from human faeces","volume":"55","author":"Domingo","year":"2005","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"326","DOI":"10.3343\/alm.2013.33.5.326","article-title":"Evaluation of vancomycin resistance 3 multiplexed pcr assay for detection of vancomycin-resistant enterococci from rectal swabs","volume":"33","author":"Cekin","year":"2013","journal-title":"Ann. Lab. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1099\/jmm.0.001159","article-title":"Rapid multiplex detection of the resistance genes mecA, vanA and vanB from Gram-positive cocci-positive blood cultures using a PCR-dipstick technique","volume":"69","author":"Shanmugakani","year":"2020","journal-title":"J. Med. Microbiol."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/11\/11\/2081\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:28:05Z","timestamp":1760167685000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/11\/11\/2081"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,10]]},"references-count":16,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["diagnostics11112081"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics11112081","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,10]]}}}