{"doi":"10.1186/s12936-020-03168-x","title":"Member species of the Anopheles gambiae complex can be misidentified as Anopheles leesoni","abstract":"<jats:title>Abstract</jats:title><jats:sec>\n                <jats:title>Background</jats:title>\n                <jats:p>Accurate <jats:italic>Anopheles</jats:italic> species identification is key for effective malaria vector control. Identification primarily depends on morphological analysis of field samples as well as molecular species-specific identifications. During an intra-laboratory assessment (proficiency testing) of the <jats:italic>Anopheles funestus</jats:italic> group multiplex PCR assay, it was noted that <jats:italic>Anopheles arabiensis</jats:italic> can be misidentified as <jats:italic>Anopheles leesoni</jats:italic>, a zoophilic member of the <jats:italic>An. funestus</jats:italic> group. The aim of this project was, therefore, to ascertain whether other members of the <jats:italic>Anopheles gambiae</jats:italic> complex can also be misidentified as <jats:italic>An. leesoni</jats:italic> when using the standard <jats:italic>An. funestus</jats:italic> multiplex PCR.</jats:p>\n              </jats:sec><jats:sec>\n                <jats:title>Methods</jats:title>\n                <jats:p>The <jats:italic>An. funestus</jats:italic> multiplex PCR was used to amplify DNA from <jats:italic>An. gambiae</jats:italic> complex specimens. These included specimens from the laboratory colonies and field samples from the Democratic Republic of Congo. Amplified DNA from these specimens, using the universal (UV) and <jats:italic>An. leesoni</jats:italic> species-specific primers (LEES), were sequence analysed. Additionally, <jats:italic>An. leesoni</jats:italic> DNA was processed through the diagnostic <jats:italic>An. gambiae</jats:italic> multiplex PCR to determine if this species can be misidentified as a member of the <jats:italic>An. gambiae</jats:italic> complex.</jats:p>\n              </jats:sec><jats:sec>\n                <jats:title>Results</jats:title>\n                <jats:p>Laboratory-colonized as well as field-collected samples of <jats:italic>An. arabiensis</jats:italic>, <jats:italic>An. gambiae</jats:italic>, <jats:italic>Anopheles merus</jats:italic>, <jats:italic>Anopheles quadriannulatus, Anopheles coluzzii</jats:italic> as well as <jats:italic>Anopheles moucheti</jats:italic> produced an amplicon of similar size to that of <jats:italic>An. leesoni</jats:italic> when using an <jats:italic>An. funestus</jats:italic> multiplex PCR. Sequence analysis confirmed that the UV and LEES primers amplify a segment of the ITS2 region of members of the <jats:italic>An. gambiae</jats:italic> complex and <jats:italic>An. moucheti</jats:italic>. The reverse was not true, i.e. the <jats:italic>An. gambiae</jats:italic> multiplex PCR does not amplify DNA from <jats:italic>An. leesoni</jats:italic>.</jats:p>\n              </jats:sec><jats:sec>\n                <jats:title>Conclusion</jats:title>\n                <jats:p>This investigation shows that <jats:italic>An. arabiensis</jats:italic>, <jats:italic>An. gambiae</jats:italic>, <jats:italic>An. merus</jats:italic>, <jats:italic>An. quadriannulatus, An. coluzzii</jats:italic> and <jats:italic>An. moucheti</jats:italic> can be misidentified as <jats:italic>An. leesoni</jats:italic> when using <jats:italic>An. funestus</jats:italic> multiplex PCR. This shows the importance of identifying specimens using standard morphological dichotomous keys as far as possible prior to the use of appropriate PCR-based identification methods. Should there be doubt concerning field-collected specimens molecularly identified as <jats:italic>An. leesoni</jats:italic>, the <jats:italic>An. gambiae</jats:italic> multiplex PCR and sequencing of the internal transcribed spacer 2 (ITS2) can be used to eliminate false identifications.</jats:p>\n              </jats:sec>","journal":"Malaria Journal","year":2020,"id":40561,"datarank":1.2385717188367957,"base_score":3.4339872044851463,"endowment":3.4339872044851463,"self_citation_contribution":0.515098080672772,"citation_network_contribution":0.7234736381640239,"self_endowment_contribution":0.515098080672772,"citer_contribution":0.7234736381640239,"corpus_percentile":null,"corpus_rank":null,"citation_count":30,"citer_count":27,"citers_with_citation_signal":18,"citers_with_endowment":18,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":196853,"name":"Allison Hendershot","orcid":null,"position":1,"is_corresponding":false},{"id":196854,"name":"Minishca Dhoogra","orcid":null,"position":2,"is_corresponding":false},{"id":196855,"name":"Henry Julius","orcid":null,"position":3,"is_corresponding":false},{"id":196856,"name":"Jacek Zawada","orcid":null,"position":4,"is_corresponding":false},{"id":196857,"name":"Maria Kaiser","orcid":null,"position":5,"is_corresponding":false},{"id":196858,"name":"Neil F. 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Koekemoer","orcid":null,"position":8,"is_corresponding":false},{"id":196852,"name":"Yael Dahan-Moss","orcid":"0000-0002-3792-9708","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":3.4339872044851463,"endowment":3.4339872044851463,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"32093677","pmcid":"PMC7038563","openalex_id":"https://openalex.org/W3012794034","authors":[],"funders":[{"funder_name":"National Institute for Communicable Diseases","grant_id":"90932","title":null},{"funder_name":"DST/NRF South African Chairs Initiative","grant_id":"64763","title":null},{"funder_name":"Eck Institute for Global Health, University of Notre Dame","grant_id":"Pilot project grant","title":null}],"total_grants":3,"fwci":3.0573,"citation_percentile":0.92001676,"influential_citations":0,"citation_trend":[{"year":2020,"count":1},{"year":2021,"count":1},{"year":2022,"count":7},{"year":2023,"count":8},{"year":2024,"count":4},{"year":2025,"count":4},{"year":2026,"count":5}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://malariajournal.biomedcentral.com/track/pdf/10.1186/s12936-020-03168-x","host_type":"journal"},{"url":"https://malariajournal.biomedcentral.com/track/pdf/10.1186/s12936-020-03168-x","host_type":"GOLD"},{"url":"https://malariajournal.biomedcentral.com/track/pdf/10.1186/s12936-020-03168-x","host_type":"publisher"},{"url":"http://link.springer.com/content/pdf/10.1186/s12936-020-03168-x.pdf","host_type":"publisher"},{"url":"http://link.springer.com/article/10.1186/s12936-020-03168-x/fulltext.html","host_type":"publisher"},{"url":"https://doi.org/10.1186/s12936-020-03168-x","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/32093677","host_type":"repository"},{"url":"https://doaj.org/article/ff87a0d88be84877a37bc81a1ab006ea","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/7038563","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC7038563","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC7038563?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["Malaria Research and Control","Parasitic Diseases Research and Treatment","Mosquito-borne diseases and control","Biology","Medicine","Environmental Science","Animals","Anopheles","DNA","Democratic Republic of the Congo","Malaria","Mosquito Vectors","Multiplex Polymerase Chain Reaction","Species Specificity"],"mesh_terms":["Mosquito Vectors","Animals","Anopheles","DNA","Malaria","Species Specificity","Democratic Republic of the Congo","Multiplex Polymerase Chain Reaction"],"keywords":["Anopheles gambiae","Anopheles","Entomology","Parasitology","Biology","Tropical medicine","Zoology","Malaria","Ecology","Immunology","Morphology","Species identification","Anopheles Leesoni","Dichotomous Keys","An. 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