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Sixty-three isolates demonstrating microbiological features typical for pneumococci (optochin susceptibility and\/or bile solubility) were investigated by phenotypic tests,<jats:italic>lytA<\/jats:italic>and 16S rRNA gene polymorphism and whole-genome sequencing (WGS). All isolates had a 6-bp deletion in the<jats:italic>lytA<\/jats:italic>3\u2032 terminus, characteristic for Mitis streptococc and all but two isolates lacked the pneumococcal signature cytosine at nucleotide position 203 in the 16S rRNA genes. The counterparts of<jats:italic>psaA<\/jats:italic>and<jats:italic>ply<\/jats:italic>were present in 100% and 81.0% of isolates, respectively; the spn9802 and spn9828 loci were characteristic for 49.2% and 38.1% of isolates, respectively. Phylogenetic trees and networks, based on the multilocus sequence analysis (MLSA) scheme, ribosomal multilocus sequence typing (rMLST) scheme and core-genome analysis, clearly separated investigated isolates from<jats:italic>S. pneumoniae<\/jats:italic>and demonstrated the polyclonal character of misID streptococci, associated with the<jats:italic>Streptococcus pseudopneumoniae<\/jats:italic>and<jats:italic>Streptococcus mitis<\/jats:italic>groups. While the<jats:italic>S. pseudopneumoniae<\/jats:italic>clade was relatively well defined in all three analyses, only the core-genome analysis revealed the presence of another cluster comprising a fraction of misID streptococci and a strain proposed elsewhere as a representative of a novel species in the Mitis group. Our findings point to complex phylogenetic and taxonomic relationships among<jats:italic>S. mitis-<\/jats:italic>like bacteria and support the notion that this group may in fact consist of several distinct species.<\/jats:p>","DOI":"10.1007\/s10096-020-03916-6","type":"journal-article","created":{"date-parts":[[2020,7,8]],"date-time":"2020-07-08T10:03:43Z","timestamp":1594202623000},"page":"1865-1878","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Relationships among streptococci from the mitis group, misidentified as Streptococcus pneumoniae"],"prefix":"10.1007","volume":"39","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0284-6440","authenticated-orcid":false,"given":"Ewa","family":"Sadowy","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Agnieszka","family":"Bojarska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicja","family":"Kuch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Skoczy\u0144ska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keith A.","family":"Jolley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin C. 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