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Here, we examined\n                    <jats:italic>E. coli<\/jats:italic>\n                    O101\/O162 O-serogroup epidemiology among bacteremia isolates and performed genetic, biochemical and structural analyses. We demonstrate that the O101\/O162 O-serogroup is globally widespread and highly associated with multidrug resistance (MDR). Dominant lineages belonged to clonal complex 10, including the high-risk clone ST167. Most O101\/O162 ExPEC isolates contained an unusual O101\n                    <jats:italic>rfb<\/jats:italic>\n                    locus previously identified as Onovel32, of which 30% harbored disruptions in the methyltransferase (MT) encoding gene. These disruptions resulted in alteration of the polysaccharide composition, and we identified two major O101 variants. Additionally, we found that an intact MT is a prerequisite for\n                    <jats:italic>O<\/jats:italic>\n                    -methylation of the polysaccharide, indicating terminal capping. Our data supports a model of a co-polymeric O101 O-antigen structure, which is unique for\n                    <jats:italic>E. coli<\/jats:italic>\n                    . Finally, we established that the two clinically relevant O101 O-antigen variant structures are highly immunogenic as polysaccharide-protein conjugates but differ in their ability to elicit opsonophagocytic antibodies. Altogether, our data provides important insights into O-serotype epidemiology and O-polysaccharide variation within\n                    <jats:italic>E. coli<\/jats:italic>\n                    O101\/O162 isolates associated with bacteremia.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-45688-7","type":"journal-article","created":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T16:04:14Z","timestamp":1774281854000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Molecular epidemiology and structural diversity of O101\/O162 O-antigen variants among Escherichia coli bacteremia isolates"],"prefix":"10.1038","volume":"16","author":[{"given":"Eveline","family":"Weerdenburg","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark","family":"de Been","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aldert","family":"Zomer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wannisa","family":"Ritmahan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joyce","family":"L\u00fcbbers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alan B.","family":"Moran","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simone","family":"Nicolardi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manfred","family":"Wuhrer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Neil","family":"Ravenscroft","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chakkumkal","family":"Anish","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeroen","family":"Geurtsen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michel","family":"Beurret","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,3,23]]},"reference":[{"key":"45688_CR1","doi-asserted-by":"publisher","first-page":"1288","DOI":"10.1017\/ice.2016.174","volume":"37","author":"LM Weiner","year":"2016","unstructured":"Weiner, L. 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EW, MdB, JL, AM and MB are employees of Janssen Vaccines & Prevention B.V. and may hold stocks or shares in Johnson & Johnson, LLC. JG and CA were employees of Janssen Vaccines & Prevention B.V. at the time of the study and may hold stocks or shares in Johnson & Johnson, LLC. JG is currently an employee of Sanofi. CA is currently an employee of AstraZeneca. Other authors declare no conflict of interest related to this work.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"The current studies are not human subject studies. During this study, only microorganisms were examined sourced from observational surveillance studies described previously\n                      6,27,28\n                      . All methods were carried out and reported in accordance with relevant guidelines and regulations. Animal studies were approved by the local IACUC committee at Johnson & Johnson in Spring House, Pennsylvania USA, under AUP: IDV510S, and reported in accordance with ARRIVE guidelines. Rats were anesthetized with CO\n                      2\n                      and euthanized by cutting the diaphragm. Animals were housed and immunization studies were completed in accordance with the Guide for the Care and Use of Animals and Animal Welfare Act in an AAALAC-accredited facility.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics considerations"}}],"article-number":"14777"}}