{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T12:58:11Z","timestamp":1792155491263,"version":"4.4.0"},"reference-count":32,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2017,12,1]],"date-time":"2017-12-01T00:00:00Z","timestamp":1512086400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"National Research Foundation of South Africa","award":["DCOE017"],"award-info":[{"award-number":["DCOE017"]}]},{"name":"Wits Enterprize"},{"name":"Technology Innovation Agency"},{"DOI":"10.13039\/100000011","name":"Howard Hughes Medical Institute","doi-asserted-by":"publisher","award":["HHMI000"],"award-info":[{"award-number":["HHMI000"]}],"id":[{"id":"10.13039\/100000011","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001322","name":"South African Medical Research Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001342","name":"Department of Science and Technology, Republic of South Africa","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001342","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2017,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Molecular diagnostics have revolutionized the management of health care through enhanced detection of disease or infection and effective enrollment into treatment. In recognition of this, the World Health Organization approved the rollout of nucleic acid amplification technologies for identification of\n            <jats:named-content content-type=\"genus-species\">Mycobacterium tuberculosis<\/jats:named-content>\n            using platforms such as GeneXpert MTB\/RIF, the GenoType MTBDR\n            <jats:italic>plus<\/jats:italic>\n            line probe assay, and, more recently, GeneXpert MTB\/RIF Ultra. These assays can simultaneously detect tuberculosis infection and assess rifampin resistance. However, their widespread use in health systems requires verification and quality assurance programs. To enable development of these, we report the construction of genetically modified strains of\n            <jats:named-content content-type=\"genus-species\">Mycobacterium smegmatis<\/jats:named-content>\n            that mimic the profile of\n            <jats:named-content content-type=\"genus-species\">Mycobacterium tuberculosis<\/jats:named-content>\n            on both the GeneXpert MTB\/RIF and the MTBDR\n            <jats:italic>plus<\/jats:italic>\n            line probe diagnostic tests. Using site-specific gene editing, we also created derivatives that faithfully mimic the diagnostic result of rifampin-resistant\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis<\/jats:named-content>\n            , with mutations at positions 513, 516, 526, 531, and 533 in the rifampin resistance-determining region of the\n            <jats:italic>rpoB<\/jats:italic>\n            gene. Next, we extended this approach to other diseases and demonstrated that a\n            <jats:named-content content-type=\"genus-species\">Staphylococcus aureus<\/jats:named-content>\n            gene sequence can be introduced into\n            <jats:named-content content-type=\"genus-species\">M. smegmatis<\/jats:named-content>\n            to generate a positive response for the SCC\n            <jats:italic>mec<\/jats:italic>\n            probe in the GeneXpert SA Nasal Complete molecular diagnostic cartridge, designed for identification of methicillin-resistant\n            <jats:named-content content-type=\"genus-species\">S. aureus<\/jats:named-content>\n            . These biomimetic strains are cost-effective, have low biohazard content, accurately mimic drug resistance, and can be produced with relative ease, thus illustrating their potential for widespread use as verification standards for diagnosis of a variety of diseases.\n          <\/jats:p>","DOI":"10.1128\/jcm.01111-17","type":"journal-article","created":{"date-parts":[[2017,9,21]],"date-time":"2017-09-21T00:30:24Z","timestamp":1505953824000},"page":"3384-3394","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":7,"title":["Genetic Mimetics of Mycobacterium tuberculosis and Methicillin-Resistant Staphylococcus aureus as Verification Standards for Molecular Diagnostics"],"prefix":"10.1128","volume":"55","author":[{"given":"Edith Erika","family":"Machowski","sequence":"first","affiliation":[{"name":"DST\/NRF Centre of Excellence for Biomedical TB Research, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bavesh Davandra","family":"Kana","sequence":"additional","affiliation":[{"name":"DST\/NRF Centre of Excellence for Biomedical TB Research, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa"},{"name":"MRC-CAPRISA HIV-TB Pathogenesis and Treatment Research Unit, Centre for the AIDS Programme of Research in South Africa, Durban, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.25956"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/npjpcrm.2016.78"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0732-8893(97)00006-0"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccm.2009.08.016"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1155\/2008\/857901"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.TBTB2-0040-2016"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-319-19132-4"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.05167-11"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/351456a0"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0152694"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/0140-6736(93)90417-F"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/emi.2014.17"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkl058"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.33073\/pjm-2011-013"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0402521101"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.385138"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.6.1549-1555.2000"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.03340-14"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0152926"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa0907847"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1164\/rccm.201103-0536OC"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01715-13"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1136\/jcp.34.8.927"},{"key":"e_1_3_2_25_2","volume-title":"Treatment guidelines for drug-resistant tuberculosis, 2016 update","author":"World Health Organization","unstructured":"World Health Organization. Treatment guidelines for drug-resistant tuberculosis, 2016 update. World Health Organization, Geneva, Switzerland."},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature05441"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.3390\/diagnostics5020200"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.84.1.166"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01884-08"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01714-08"},{"key":"e_1_3_2_31_2","first-page":"77","article-title":"Gene replacement using pretreated DNA","volume":"54","author":"Gordhan BG","year":"2001","unstructured":"Gordhan BG Parish T . 2001. Gene replacement using pretreated DNA. 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