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Since its initial detection in Djibouti in 2012, malaria morbidity and mortality have increased exponentially in the country. With the observed increases in human malaria cases and previously reported An. stephensi involvement in outbreaks, high-quality evidence of Plasmodium sporozoite-carrying\n                      <jats:italic>An.<\/jats:italic>\n                      <jats:italic>stephensi<\/jats:italic>\n                      is essential to confirm and monitor the role of the invasive vector in malaria transmission dynamics in Djibouti. This study aims to detect Plasmodium sporozoites in\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      collected in Djibouti using a multiplex circumsporozoite ELISA bead assay and assess the genetic relatedness of these populations to\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      across the Horn of Africa.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      One hundred and ninety-six wild caught adult\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      mosquitoes from Djibouti City were collected, morphologically identified, and molecularly confirmed using a portion of the cytochrome c oxidase subunit 1 marker. All samples were then tested for infective sporozoites\n                      <jats:italic>Plasmodium vivax 210, P. vivax 247 and P. falciparum<\/jats:italic>\n                      using a multiplex circumsporozoite enzyme-linked immunosorbent assay bead assay. One hundred and fourteen COI sequences that passed further sequence analysis quality control for comparative population genetic analysis were used to characterize the genetic diversity of the sampled\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      and its genetic relatedness to other\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      populations across the Horn of Africa.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      All 196 samples were morphologically and molecularly confirmed to be\n                      <jats:italic>An. stephensi.<\/jats:italic>\n                      Plasmodium vivax 210 sporozoites were detected in two specimens with a positivity rate of 1.0%. No Pv247 or Pf sporozoites were detected. An analysis of the COI region showed that the Pv210-infected\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      belonged to the dominant invasive haplotypes circulating in the Horn of Africa.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      The findings from this study confirm the involvement of\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      in P. vivax transmission in Djibouti and the genetic relatedness of Djiboutian\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      populations to other populations across the Horn of Africa. This highlights the threat of\n                      <jats:italic>An. stephensi<\/jats:italic>\n                      invasion and supports a rapid and comprehensive response, particularly through surveillance and control.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-026-07532-9","type":"journal-article","created":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T18:44:05Z","timestamp":1783536245000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Evidence of Anopheles stephensi involvement in the transmission of Plasmodium vivax in Djibouti City using highly sensitive sporozoite detection assay"],"prefix":"10.1186","volume":"19","author":[{"given":"Subhika","family":"Rao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2391-6583","authenticated-orcid":false,"given":"Jeanne N.","family":"Samake","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Rafferty","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Mumba","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheleme","family":"Chibsa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meshesha","family":"Balkew","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bouh Abdi","family":"Khaireh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samatar Kayad","family":"Guelleh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed Mousse","family":"Ibrahim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdoulilah Ahmed","family":"Abdi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Zohdy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,7,8]]},"reference":[{"key":"7532_CR1","unstructured":"WHO. 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