{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T07:49:08Z","timestamp":1770968948490,"version":"3.50.1"},"reference-count":30,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2011,1,5]]},"abstract":"<jats:p>Malaria and filariasis are transmitted in the Southwest Pacific region by <jats:italic>Anopheles punctulatus<\/jats:italic> sibling species including <jats:italic>An. punctulatus<\/jats:italic>, <jats:italic>An. koliensis<\/jats:italic>, the <jats:italic>An. farauti<\/jats:italic> complex 1\u20138 (includes <jats:italic>An. hinesorum<\/jats:italic> [<jats:italic>An. farauti<\/jats:italic> 2], <jats:italic>An. torresiensis<\/jats:italic> [<jats:italic>An. farauti<\/jats:italic> 3]). Distinguishing these species from each other requires molecular diagnostic methods. We developed a multiplex polymerase chain reaction (PCR)\u2013based assay specific for known species-specific nucleotide differences in the internal transcribed spacer 2 region and identified the five species most frequently implicated in transmitting disease (<jats:italic>An. punctulatus<\/jats:italic>, <jats:italic>An. koliensis<\/jats:italic>, <jats:italic>An. farauti<\/jats:italic> 1, <jats:italic>An. hinesorum<\/jats:italic>, and <jats:italic>An. farauti<\/jats:italic> 4). A set of 340 individual mosquitoes obtained from seven Papua New Guinea provinces representing a variety of habitats were analyzed by using this multiplex assay. Concordance between molecular and morphological diagnosis was 56.4% for <jats:italic>An. punctulatus<\/jats:italic>, 85.3% for <jats:italic>An. koliensis<\/jats:italic>, and 88.9% for <jats:italic>An. farauti<\/jats:italic>. Among 158 mosquitoes morphologically designated as <jats:italic>An. farauti<\/jats:italic>, 33 were re-classified by PCR as <jats:italic>An. punctulatus<\/jats:italic>, 4 as <jats:italic>An. koliensis<\/jats:italic>, 26 as <jats:italic>An. farauti<\/jats:italic> 1, 49 as <jats:italic>An. hinesorum<\/jats:italic>, and 46 as <jats:italic>An. farauti<\/jats:italic> 4. Misclassification results from variable coloration of the proboscis and overlap of <jats:italic>An. punctulatus<\/jats:italic>, <jats:italic>An. koliensis<\/jats:italic>, the <jats:italic>An. farauti<\/jats:italic> 4. This multiplex technology enables further mosquito strain identification and simultaneous detection of microbial pathogens.<\/jats:p>","DOI":"10.4269\/ajtmh.2011.10-0438","type":"journal-article","created":{"date-parts":[[2011,1,6]],"date-time":"2011-01-06T16:27:08Z","timestamp":1294331228000},"page":"166-173","source":"Crossref","is-referenced-by-count":17,"title":["High Throughput Multiplex Assay for Species Identification of Papua New Guinea Malaria Vectors: Members of the Anopheles punctulatus (Diptera: Culicidae) Species Group"],"prefix":"10.4269","volume":"84","author":[{"given":"Cara N.","family":"Henry-Halldin","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lisa","family":"Reimer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Edward","family":"Thomsen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gussy","family":"Koimbu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Allison","family":"Zimmerman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John B.","family":"Keven","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henry","family":"Dagoro","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manuel W.","family":"Hetzel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ivo","family":"Mueller","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Siba","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter A.","family":"Zimmerman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2795","reference":[{"key":"R1","series-title":"J Parasitol","first-page":"95","article-title":"The punctulatus complex of Anopheles (Diptera: Culicidae)","volume":"32","author":"Rozeboom","year":"1946"},{"key":"R2","series-title":"Trans R Entomol Soc Lond","first-page":"413","article-title":"Morphological studies on the Anopheles punctulatus D\u00f6nitz complex","volume":"125","author":"Bryan","year":"1974"},{"key":"R3","series-title":"Bull Entomol Res","first-page":"463","article-title":"Blood-feeding and resting behaviour in the Anopheles punctulatus D\u00f6nitz complex (Diptera: Culicidae) from coastal Papua New Guinea","volume":"75","author":"Charlwood","year":"1985"},{"key":"R4","series-title":"J Med Entomol","first-page":"16","article-title":"Speciation and distribution of the members of the Anopheles punctulatus (Diptera: Culicidae) group in Papua New Guinea","volume":"39","author":"Cooper","year":"2002"},{"key":"R5","series-title":"Med Vet Entomol","first-page":"37","article-title":"Allozyme analysis reveals six species within the Anopheles punctulatus complex of mosquitoes in Papua New Guinea","volume":"7","author":"Foley","year":"1993"},{"key":"R6","series-title":"Trans R Soc Trop Med Hyg","first-page":"85","article-title":"Studies on the Anopheles punctulatus complex. 3. 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