{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:07:13Z","timestamp":1785424033082,"version":"3.56.0"},"reference-count":34,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T00:00:00Z","timestamp":1775520000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T00:00:00Z","timestamp":1775520000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"National Science Centre (NCN) \"SONATA\"","award":["2021\/43\/D\/NZ6\/01250"],"award-info":[{"award-number":["2021\/43\/D\/NZ6\/01250"]}]},{"name":"\"The Excellence Initiative - Research University\" (IDUB) programme","award":["BOB-IDUB-622-616\/2023"],"award-info":[{"award-number":["BOB-IDUB-622-616\/2023"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,4,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Currently, the\n                      <jats:italic toggle=\"yes\">Mycobacterium kansasii<\/jats:italic>\n                      complex (MKC) species identification involves PCR amplification, followed by sequencing or digestion of the amplicons. The purpose of this study was to develop a one-step multiplex PCR assay allowing for a fast and accurate identification of MKC species. A total of 158\n                      <jats:italic toggle=\"yes\">Mycobacterium<\/jats:italic>\n                      sp. genomes were searched for sites that would yield easily detectable amplicons of different sizes among the MKC species while producing no amplicons for other-than-MKC mycobacterial species. Three primer sets were designed and tested\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      on seven reference strains, representing all MKC species. For the evaluation of the method, a total of 136\n                      <jats:italic toggle=\"yes\">Mycobacterium<\/jats:italic>\n                      sp. isolates were analyzed. The vast majority (96\/98, 98%) of the tested MKC strains yielded a species-characteristic profile, consistent with\n                      <jats:italic toggle=\"yes\">in silico<\/jats:italic>\n                      predictions. The only two isolates, which gave incongruent results, belong to atypical\n                      <jats:italic toggle=\"yes\">M. kansasii<\/jats:italic>\n                      subtype IIB and were designated as\n                      <jats:italic toggle=\"yes\">M. persicum<\/jats:italic>\n                      with a newly designed assay. No products were obtained for mycobacteria other than MKC. This study offers a new PCR-based method for identification of all MKC species. It involves a single-step protocol and yields reproducible and easily interpretable results.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      Current methods used for the\n                      <jats:italic toggle=\"yes\">Mycobacterium kansasii<\/jats:italic>\n                      complex (MKC) species identification lack the resolution to differentiate between individual MKC species and require time-demanding multiple steps and costly analyses. Herein, we present a novel one-step multiplex PCR assay that enables rapid and cost-effective species-level identification of MKC members, suitable for implementation in both clinical diagnostics and epidemiological investigations. The method was evaluated using 143\n                      <jats:italic toggle=\"yes\">Mycobacterium<\/jats:italic>\n                      strains, including 17 reference strains and 126 clinical or environmental isolates. It accurately identified the vast majority (98%) of strains, with only two atypical\n                      <jats:italic toggle=\"yes\">M. kansasii<\/jats:italic>\n                      subtype IIB isolates misidentified as\n                      <jats:italic toggle=\"yes\">M. persicum<\/jats:italic>\n                      . The calculated sensitivity and specificity of the assay were 98% and 100%, respectively.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03267-25","type":"journal-article","created":{"date-parts":[[2026,3,5]],"date-time":"2026-03-05T14:00:41Z","timestamp":1772719241000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["One-step multiplex PCR assay for identification of\n                    <i>Mycobacterium kansasii<\/i>\n                    complex species"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6165-8330","authenticated-orcid":true,"given":"Paulina Joanna","family":"Wa\u0142puska","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00yd0p282","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Microbiology, Institute of Microbiology, Faculty of Biology, University of 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