{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T13:40:07Z","timestamp":1792676407232,"version":"4.5.0"},"reference-count":43,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T00:00:00Z","timestamp":1792627200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T00:00:00Z","timestamp":1792627200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R21AI176642"],"award-info":[{"award-number":["R21AI176642"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R00 AI153483"],"award-info":[{"award-number":["R00 AI153483"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mSphere"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Klebsiella pneumoniae<\/jats:italic>\n                      and related species are a common cause of healthcare-associated infections. The gut is a major\n                      <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                      reservoir, and gut colonization is a risk factor for developing an extraintestinal\n                      <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                      infection. Patients can be colonized by multiple\n                      <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                      strains or even species in the gut simultaneously, and there is high concordance between the gut-colonizing and infection-causing strains. The detection and characterization of colonizing strains is critical for a better understanding of the progression to infection and for developing interventions for colonized patients. However, the association between mixed or mono-colonization and subsequent infection is unknown. In this study, we developed an amplicon-based sequencing method called\n                      <jats:italic toggle=\"yes\">wzi<\/jats:italic>\n                      -Seq that enables the detection and quantification of\n                      <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                      strains from complex samples and mixtures using the conserved capsule gene\n                      <jats:italic toggle=\"yes\">wzi<\/jats:italic>\n                      as a molecular barcode. This method is highly accurate and precise, with a sensitivity of 93% and specificity of 99.8% in mixtures containing as many as 58 unique\n                      <jats:italic toggle=\"yes\">wzi<\/jats:italic>\n                      types, and 90.8% concordance with culture-based detection from archived rectal swab samples from an established case and control cohort. Adjudicating\n                      <jats:italic toggle=\"yes\">wzi<\/jats:italic>\n                      -Seq with culture data, we determined that 63.2% (108\/171) patients were mono-colonized with a single\n                      <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                      strain, while 36.8% (63\/171) had mixed colonization with multiple\n                      <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                      strains. Controlling for patient variables in multivariate analysis, we determined that mono-colonization was significantly (\n                      <jats:italic toggle=\"yes\">P<\/jats:italic>\n                      = 0.034) associated with infection. Characterization of\n                      <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                      colonizing populations could improve the accuracy of assessing infection risk and enable targeted interventions to prevent these healthcare-associated infections.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        gut colonization is a major risk factor for the development of extraintestinal\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        infections in hospitalized settings. However, it is unknown if patients are colonized by one or multiple strains of\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        and how the population structure of\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        in the gut impacts infection risk. Here, we describe the development of a novel technique,\n                        <jats:italic toggle=\"yes\">wzi<\/jats:italic>\n                        -Seq, to detect and quantify multiple\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        strains from complex samples using standard techniques and rapid DNA sequencing. We applied\n                        <jats:italic toggle=\"yes\">wzi<\/jats:italic>\n                        -Seq to rectal swabs from a well-characterized patient cohort and found that\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        colonization with a single strain was more prevalent than colonization with multiple strains. Furthermore, mono-colonized patients had a significantly higher risk of developing a\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        infection than mixed-colonized patients. This work validates a new tool to study\n                        <jats:italic toggle=\"yes\">Klebsiella<\/jats:italic>\n                        populations and reveals that population structure in the gut influences the risk of healthcare-associated infections.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/msphere.00329-26","type":"journal-article","created":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T13:00:41Z","timestamp":1792674041000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["A novel technique to characterize\n                    <i>Klebsiella pneumoniae<\/i>\n                    populations indicates that mono-colonization is associated with risk of infection"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3618-9194","authenticated-orcid":true,"given":"Lavinia 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