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Despite advancements in detection methods like ultrasound and laparoscopy, these techniques remain limited by low specificity and invasiveness, underscoring the need for a highly specific, noninvasive in\u00a0vitro diagnostic method. This study investigates the potential of using menstrual blood as a noninvasive diagnostic sample for endometriosis by targeting genetic and inflammatory markers associated with endometriosis lesions. A novel digital droplet enzyme-linked immunosorbent assay (ddELISA) was developed, leveraging SiO\n            <jats:sub>2<\/jats:sub>\n            nanoparticles for the femtomolar-sensitive detection of inflammatory cytokines (OPN, IL-10, IL-6) in menstrual blood. Single-cell RNA sequencing revealed differentiation patterns across endometrial tissues and menstrual blood, affirming that menstrual blood replicates key inflammatory and immune properties of endometriosis. Furthermore, endometriosis menstrual blood endometrial cells derived from human menstrual blood displayed similar properties to endometrial stromal cells in endometriosis lesions, validating menstrual blood as a suitable in\u00a0vitro diagnostic sample. In contrast to traditional ELISA, ddELISA supports multi-target detection with enhanced sensitivity and reduced processing time, allowing precise biomarker analysis from minimal sample volumes. Our ddELISA-based approach shows promise as a rapid, accessible, and accurate diagnostic tool for endometriosis, with potential for practical clinical application.\n          <\/jats:p>","DOI":"10.34133\/research.0652","type":"journal-article","created":{"date-parts":[[2025,3,13]],"date-time":"2025-03-13T16:53:11Z","timestamp":1741884791000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark_01","source":"Crossref","is-referenced-by-count":11,"title":["A Noninvasive Menstrual Blood-Based Diagnostic Platform for Endometriosis Using Digital Droplet Enzyme-Linked Immunosorbent Assay and Single-Cell RNA Sequencing"],"prefix":"10.34133","volume":"8","author":[{"given":"Han","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, The Second Affiliated Hospital and Yuying Children\u2019s Hospital of Wenzhou Medical University, Wenzhou","place":["China"]},{"name":"Oncology Discipline Group, \rThe Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhouyi","family":"Gan","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, The Second Affiliated Hospital and Yuying Children\u2019s Hospital of Wenzhou Medical University, Wenzhou","place":["China"]},{"name":"Oncology Discipline Group, \rThe Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yueyue","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, The Second Affiliated Hospital and Yuying Children\u2019s Hospital of Wenzhou Medical University, Wenzhou","place":["China"]},{"name":"Oncology Discipline Group, \rThe Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dingmeng","family":"Hu","sequence":"additional","affiliation":[{"name":"Joint Centre of Translational Medicine, \rThe First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325035, China."},{"name":"Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision, and Brain Health), Wenzhou Institute, \rUniversity of Chinese Academy of Sciences, Wenzhou, Zhejiang 325000, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lexiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Joint Centre of Translational Medicine, \rThe First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325035, China."},{"name":"Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision, and Brain Health), Wenzhou Institute, \rUniversity of Chinese Academy of Sciences, Wenzhou, Zhejiang 325000, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangfu","family":"Ye","sequence":"additional","affiliation":[{"name":"Joint Centre of Translational Medicine, \rThe First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325035, China."},{"name":"Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision, and Brain Health), Wenzhou Institute, \rUniversity of Chinese Academy of Sciences, Wenzhou, Zhejiang 325000, China."},{"name":"Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, \rChinese Academy of Sciences, Beijing 100190, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0006-8950","authenticated-orcid":true,"given":"Ping","family":"Duan","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, The Second Affiliated Hospital and Yuying Children\u2019s Hospital of Wenzhou Medical University, Wenzhou","place":["China"]},{"name":"Oncology Discipline Group, \rThe Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","published-online":{"date-parts":[[2025,4]]},"reference":[{"issue":"4","key":"e_1_3_4_2_2","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1038\/s41574-024-00959-z","article-title":"Targeting endometriosis","volume":"20","author":"Greenhill C","year":"2024","unstructured":"Greenhill C. 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