{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,14]],"date-time":"2026-03-14T06:43:10Z","timestamp":1773470590230,"version":"3.50.1"},"reference-count":37,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,1,7]],"date-time":"2022-01-07T00:00:00Z","timestamp":1641513600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002855","name":"Ministry of Science and Technology of the People's Republic of China","doi-asserted-by":"publisher","award":["2017YFC1601400"],"award-info":[{"award-number":["2017YFC1601400"]}],"id":[{"id":"10.13039\/501100002855","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>The aim of this study was to develop homogeneous and stable plasmid DNA reference materials for detecting the mechanisms of resistance to quinolones and fluoroquinolones in foodborne pathogens. The DNA fragments of 11 target genes associated with quinolone and fluoroquinolone resistance were artificially synthesized, inserted into plasmid vectors, and transferred into recipient cells. PCR and sequencing of DNA were performed to assess the genetic stability of the target DNA in recombinant Escherichia coli DH5\u03b1 cells during subculturing for 15 generations. The limit of detection (LOD) of the target DNA was determined using PCR and real-time qualitative PCR (qPCR). The homogeneity and storage stability of plasmid DNA reference materials were evaluated in terms of plasmid DNA quantity, PCR-measured gene expression, and qPCR threshold cycle. All 11 target DNAs were successfully synthesized and inserted into vectors to obtain recombinant plasmids. No nucleotide mutations were identified in the target DNA being stably inherited and detectable in the corresponding plasmids during subculturing of recombinant strains. When the target DNA was assessed using PCR and qPCR, the LOD was \u22641.77 \u00d7 105 and 3.26 \u00d7 104 copies\/\u03bcL, respectively. Further, when the reference materials were stored at 37 \u00b0C for 13 days, 4 \u00b0C for 90 days, and \u221220 \u00b0C for 300 days, each target DNA was detectable by PCR, and no mutations were found. Although the threshold cycle values of qPCR varied with storage time, they were above the LOD, and no significant differences were found in the quantity of each plasmid DNA at different timepoints. Further, the homogeneity and stability of the materials were highly consistent with the requirements of standard reference materials. To summarize, considering that our plasmid DNA reference materials conformed to standard requirements, they can be used to detect the mechanisms of quinolone and fluoroquinolone resistance in foodborne pathogens.<\/jats:p>","DOI":"10.3390\/foods11020154","type":"journal-article","created":{"date-parts":[[2022,1,9]],"date-time":"2022-01-09T20:29:26Z","timestamp":1641760166000},"page":"154","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Developing Qualitative Plasmid DNA Reference Materials to Detect Mechanisms of Quinolone and Fluoroquinolone Resistance in Foodborne Pathogens"],"prefix":"10.3390","volume":"11","author":[{"given":"Qinya","family":"Niu","sequence":"first","affiliation":[{"name":"College of Food Science and Engineering, Northwest A&F University, Xianyang 712100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiumin","family":"Su","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Northwest A&F University, Xianyang 712100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luxin","family":"Lian","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Northwest A&F University, Xianyang 712100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinling","family":"Huang","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Northwest A&F University, Xianyang 712100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shutong","family":"Xue","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Northwest A&F University, Xianyang 712100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1763-714X","authenticated-orcid":false,"given":"Wei","family":"Zhou","sequence":"additional","affiliation":[{"name":"Hebei Food Inspection and Research Institute, Hebei Food Safety Key Laboratory, Shijiazhuang 050091, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongyang","family":"Zhao","sequence":"additional","affiliation":[{"name":"Chinese Academy of Inspection and Quarantine, Beijing 100176, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xing\u2019an","family":"Lu","sequence":"additional","affiliation":[{"name":"Chinese Academy of Inspection and Quarantine, Beijing 100176, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shenghui","family":"Cui","sequence":"additional","affiliation":[{"name":"National Institutes for Food and Drug Control, Beijing 100050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jia","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Chemical Technology, Shijiazhuang University, Shijiazhuang 050035, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Baowei","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Northwest A&F University, Xianyang 712100, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,7]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2020, November 06). 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