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To explore the modular structure and predictive potential of these accessory elements, presence\/absence data from 77 strains (70 accessory features) were analyzed. Methods included clustering using Euclidean and Jaccard distances, gene\u2010to\u2010gene network construction with community detection, Fisher's exact tests for associations between plasmid and virulence or antimicrobial resistance genes (AMR), random forest modeling to predict virulence labels and toxin presence (excluding direct toxin markers), and PCA for visualization. Both clustering approaches revealed broad groupings, though Jaccard clustering better captured co\u2010occurring gene patterns. The co\u2010occurrence network identified 12 modules, including a prominent plasmid\u2013virulence module centered on\n                    <jats:italic>IncF<\/jats:italic>\n                    replicons,\n                    <jats:italic>stx2<\/jats:italic>\n                    ,\n                    <jats:italic>ehxA<\/jats:italic>\n                    ,\n                    <jats:italic>toxB<\/jats:italic>\n                    , and\n                    <jats:italic>espP<\/jats:italic>\n                    . Fisher's tests showed significant associations, notably between\n                    <jats:italic>IncFIA<\/jats:italic>\n                    and\n                    <jats:italic>stx2c<\/jats:italic>\n                    (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20095.3\u2009\u00d7\u200910\n                    <jats:sup>\u22124<\/jats:sup>\n                    ). The Random Forest classifier achieved a cross\u2010validated AUC of 0.853\u2009\u00b1\u20090.067, with\n                    <jats:italic>gad<\/jats:italic>\n                    ,\n                    <jats:italic>espF<\/jats:italic>\n                    ,\n                    <jats:italic>IncFIA<\/jats:italic>\n                    , and\n                    <jats:italic>ehxA<\/jats:italic>\n                    as key predictors. PCA explained 31.1%, 12.9%, and 10.0% of the variance across the first three components, separating plasmid\u2013virulence module carriers from others. These findings indicate that a modular accessory genome structure contributes to the diversity of O157:H7.\n                    <jats:italic>IncF<\/jats:italic>\n                    plasmids and associated effectors form a highly interconnected subnetwork, and accessory markers independent of direct toxin genes can effectively predict virulence status, offering potential for rapid genomic surveillance of Shiga toxin\u2010producing\n                    <jats:italic>E. coli<\/jats:italic>\n                    (STEC).\n                  <\/jats:p>","DOI":"10.1002\/mbo3.70323","type":"journal-article","created":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T06:59:41Z","timestamp":1781161181000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Integrative Modular, Network\u2010Based, and Machine Learning Framework for Predicting Accessory Genome Functions and Virulence in\n                    <i>Escherichia coli<\/i>\n                    O157:H7"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8002-6697","authenticated-orcid":false,"given":"Sydney Menzeko","family":"Gambushe","sequence":"first","affiliation":[{"name":"Discipline of Genetics, School of Agriculture and Science, College of Agriculture, Engineering and Science University of KwaZulu\u2010Natal Durban South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oliver Tendayi","family":"Zishiri","sequence":"additional","affiliation":[{"name":"Discipline of Genetics, School of Agriculture and Science, College of Agriculture, Engineering and Science University of KwaZulu\u2010Natal Durban South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,6,10]]},"reference":[{"key":"e_1_2_11_2_1","first-page":"757","article-title":"Prevalence and Potential Link Between Escherichia coli O157:H7 Isolated From Drinking Water and Stool Samples of Diarrhoeic Children in the Amathole District, South Africa","volume":"35","author":"Abongo S. 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