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Diagnosis by light microscopy is challenging, leading to significant underdiagnosis. Two genotypes (1 and 2) have been described, with genotype 1 being the most frequently detected worldwide. As no molecularly characterized isolates from Spain have been reported, we performed a molecular characterization and evaluated sociodemographic factors of\n                      <jats:italic>D. fragilis<\/jats:italic>\n                      in patients coinfected with\n                      <jats:italic>Blastocystis<\/jats:italic>\n                      sp. and\/or\n                      <jats:italic>Giardia duodenalis<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Using a multiplex qPCR assay (Allplex\u2122 GI-Parasite Assay, Seegene), we analyzed 354 stool samples previously identified as positive for\n                      <jats:italic>Blastocystis<\/jats:italic>\n                      sp. (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u2009276),\n                      <jats:italic>G. duodenalis<\/jats:italic>\n                      (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200963), or both (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200915) by light microscopy. Eligible positive samples were selected for conventional PCR targeting the\n                      <jats:italic>SSU<\/jats:italic>\n                      rRNA gene and subsequent Sanger sequencing.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      <jats:italic>D. fragilis<\/jats:italic>\n                      was detected in 34.5% of samples by qPCR; 32.3% of\n                      <jats:italic>Blastocystis<\/jats:italic>\n                      -positive, 44.9% of\n                      <jats:italic>Giardia<\/jats:italic>\n                      -positive and 46.7% of coinfected samples. We successfully sequenced 22 isolates, all corresponding to genotype 1, identifying two distinct intra-genotypic sequence variants. Younger age was positively associated with\n                      <jats:italic>D. fragilis<\/jats:italic>\n                      , with individuals coinfected with\n                      <jats:italic>Blastocystis<\/jats:italic>\n                      sp. being markedly younger than those monoinfected. Patients with\n                      <jats:italic>G. duodenalis<\/jats:italic>\n                      and\n                      <jats:italic>D. fragilis<\/jats:italic>\n                      were mainly under 15 years of age.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      This study provides the first genotyping of\n                      <jats:italic>D. fragilis<\/jats:italic>\n                      in Spain, demonstrating the circulation of two genotype 1 variants. Findings highlight the underdiagnosis of\n                      <jats:italic>D. fragilis<\/jats:italic>\n                      and its high prevalence in young patients with other fecal-orally transmitted protists. Evaluating coinfections is crucial to understanding parasite relationships and should be considered for clinical management, especially when symptoms persist.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s10096-026-05586-2","type":"journal-article","created":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T02:54:00Z","timestamp":1783565640000},"page":"3181-3191","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Molecular epidemiology of Dientamoeba fragilis coinfections with Blastocystis sp. and Giardia duodenalis in symptomatic patients from Valencia, Spain"],"prefix":"10.1007","volume":"45","author":[{"given":"Marta","family":"Garc\u00eda-Hita","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jorge","family":"Ferriz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susana","family":"Cifre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela","family":"Tapia-Veloz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Araceli","family":"Molina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Carmena","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5977-3337","authenticated-orcid":false,"given":"Maria","family":"Trelis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,7,9]]},"reference":[{"issue":"3","key":"5586_CR1","doi-asserted-by":"publisher","first-page":"352","DOI":"10.1017\/S0031182000003929","volume":"10","author":"MW Jepps","year":"1918","unstructured":"Jepps MW, Dobell C (1918) Dientamoeba fragilis n. g., n. sp., a new Intestinal Amoeba from Man. 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