{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T07:02:36Z","timestamp":1780038156146,"version":"3.53.1"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T00:00:00Z","timestamp":1780012800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p>\n                    Amebiasis, caused by\n                    <jats:italic>Entamoeba histolytica<\/jats:italic>\n                    , remains a major public health challenge in endemic regions and is often misdiagnosed due to its morphological similarity to the non-pathogenic species\n                    <jats:italic>E. dispar<\/jats:italic>\n                    . The limitations of light microscopy and the high cost of immunological assays. This study aimed to standardize RNA extraction methods in\n                    <jats:italic>Entamoeba<\/jats:italic>\n                    to support high-quality transcriptomic analyses and molecular investigations Six RNA extraction protocols were compared using\n                    <jats:italic>E. histolytica<\/jats:italic>\n                    and\n                    <jats:italic>E. dispar<\/jats:italic>\n                    trophozoites, and RNA yield, purity, and integrity were assessed by spectrophotometry (NanoDrop), fluorometry (Qubit), and automated electrophoresis (TapeStation). The TRIzol + RNeasy protocol provided the highest purity ratios (260\/280 and 260\/230) and was selected as the standard method. Extracted RNA showed good integrity (RINe 7.2 for\n                    <jats:italic>E. histolytica<\/jats:italic>\n                    and 6.6 for\n                    <jats:italic>E. dispar<\/jats:italic>\n                    ), supporting RNA-Seq library construction. Two library preparation strategies were evaluated: poly(A) selection and ribosomal RNA (rRNA) depletion. Both generated libraries with appropriate profiles; however, poly(A) selection was more efficient, yielding higher RNA concentrations and low residual rRNA (&amp;lt;3.5%), whereas rRNA depletion remained inefficient (~87% rRNA). This standardized RNA extraction workflow combined with poly(A) selection represents an effective strategy for transcriptomic analyses in\n                    <jats:italic>Entamoeba<\/jats:italic>\n                    , providing a robust methodological foundation to advance transcriptomic studies, biomarker discovery, and a deeper understanding of\n                    <jats:italic>Entamoeba<\/jats:italic>\n                    biology.\n                  <\/jats:p>","DOI":"10.3389\/fcimb.2026.1797458","type":"journal-article","created":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T06:11:36Z","timestamp":1780035096000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Standardized RNA extraction protocol for Entamoeba species: advancing molecular diagnostics and amebiasis control"],"prefix":"10.3389","volume":"16","author":[{"given":"Gabriele","family":"Cox","sequence":"first","affiliation":[{"name":"Laboratory 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RPT01J, Fiocruz Technological Platforms Network (RPT), Vice Presidency of Research and Biological Collections (VPPCB)","place":["Rio de Janeiro, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beatriz","family":"M\u00fcller","sequence":"additional","affiliation":[{"name":"Laboratory of Applied Genomics and Bioinnovations, Oswaldo Cruz Institute","place":["Rio de Janeiro, Brazil"]},{"name":"Next-Generation Sequencing Platform \u2013 RPT01J, Fiocruz Technological Platforms Network (RPT), Vice Presidency of Research and Biological Collections (VPPCB)","place":["Rio de Janeiro, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre dos","family":"Santos","sequence":"additional","affiliation":[{"name":"Laboratory of Applied Genomics and Bioinnovations, Oswaldo Cruz Institute","place":["Rio de Janeiro, Brazil"]},{"name":"Next-Generation Sequencing Platform \u2013 RPT01J, Fiocruz Technological Platforms Network (RPT), Vice Presidency of 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Oswaldo Cruz Institute","place":["Rio de Janeiro, Brazil"]},{"name":"Bioinformatics Platform \u2013 RPT04A (VPPCB)","place":["Rio de Janeiro, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thiago","family":"Parente","sequence":"additional","affiliation":[{"name":"Laboratory of Applied Genomics and Bioinnovations, Oswaldo Cruz Institute","place":["Rio de Janeiro, Brazil"]},{"name":"Bioinformatics Platform \u2013 RPT04A (VPPCB)","place":["Rio de Janeiro, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wim","family":"Degrave","sequence":"additional","affiliation":[{"name":"Laboratory of Applied Genomics and Bioinnovations, Oswaldo Cruz Institute","place":["Rio de Janeiro, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helena","family":"Toma","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Diagnosis and Hematology, Federal University of Rio de Janeiro","place":["Rio de Janeiro, 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