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Co-infection by these organisms may influence disease manifestation and host immune response. This study aimed to determine the prevalence and molecular characterization of\n                      <jats:italic>Entamoeba<\/jats:italic>\n                      species and\n                      <jats:italic>Helicobacter pylori<\/jats:italic>\n                      among patients with gastrointestinal symptoms attending Kasr El-Aini hospital. It also investigated the association of tumor necrosis factor-alpha (TNF-\u03b1) gene polymorphisms at positions\u2009\u2212\u20091031\u00a0T\/C and\u2009\u2212\u2009308G\/A with susceptibility or protection against these infections.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      A total of 372 stool samples were collected and classified into four groups:\n                      <jats:italic>Helicobacter pylori<\/jats:italic>\n                      -only,\n                      <jats:italic>Entamoeba<\/jats:italic>\n                      -only, co-infected, and control. Controls were recruited from the same clinical setting and confirmed negative for both\n                      <jats:italic>Helicobacter pylori<\/jats:italic>\n                      and\n                      <jats:italic>Entamoeba histolytica<\/jats:italic>\n                      by molecular assays. Microscopic examination was first performed for parasitic detection, followed by DNA extraction and polymerase chain reaction (PCR) for species identification of\n                      <jats:italic>Entamoeba dispar<\/jats:italic>\n                      ,\n                      <jats:italic>Entamoeba moshkovskii<\/jats:italic>\n                      ,\n                      <jats:italic>Entamoeba histolytica<\/jats:italic>\n                      , and\n                      <jats:italic>Helicobacter pylori<\/jats:italic>\n                      . Among 93 samples positive for the\n                      <jats:italic>Entamoeba<\/jats:italic>\n                      complex,\n                      <jats:italic>Entamoeba moshkovskii<\/jats:italic>\n                      was the predominant species (53.8%), followed by\n                      <jats:italic>Entamoeba. dispar<\/jats:italic>\n                      (29.0%) and\n                      <jats:italic>Entamoeba histolytica<\/jats:italic>\n                      (17.2%). Mixed infections were observed in 19 cases. Sociodemographic factors, including age, sex, residence, water source, and animal contact, were significantly associated with the presence of\n                      <jats:italic>Entamoeba<\/jats:italic>\n                      species. Among 93\n                      <jats:italic>Helicobacter pylori<\/jats:italic>\n                      -positive participants, 44.1% carried the cytotoxin-associated gene A (CagA) virulence factor. Sex and tap water consumption showed significant associations with\n                      <jats:italic>Helicobacter pylori<\/jats:italic>\n                      infection and co-infection. Genotyping analysis revealed that the TNF-\u03b1\u2009\u2212\u20091031\u00a0T\/C TT genotype and T allele were more frequent in healthy controls, suggesting a protective role, while no significant difference was observed for the\u2009\u2212\u2009308G\/A polymorphism.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      This study highlights a notable prevalence of\n                      <jats:italic>Entamoeba moshkovskii<\/jats:italic>\n                      among the\n                      <jats:italic>Entamoeba complex<\/jats:italic>\n                      and a considerable rate of co-infection with\n                      <jats:italic>Helicobacter pylori<\/jats:italic>\n                      . Environmental and host factors, particularly water source and sex, play important roles in infection dynamics. The TNF-\u03b1\u2009\u2212\u20091031C allele appears to confer protection against gastrointestinal infection. These findings emphasize the need for improved diagnostic screening and public health measures to limit transmission and co-infection in endemic areas.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12866-026-04964-2","type":"journal-article","created":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T08:39:47Z","timestamp":1776328787000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Role of TNF-\u03b1 genetic variants in coinfection with Helicobacter pylori and the Entamoeba complex: a cross-sectional study"],"prefix":"10.1186","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8119-9549","authenticated-orcid":false,"given":"Asmaa","family":"Ibrahim","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moamen A. 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