A multiplex PCR assay for the identification of 5 sibling species of the Lindesayi Complex of the genus Anopheles (Diptera: Culicidae) in Bhutan

Li Zhou; Rinsin Namgay; Atiporn Saeung; Jassada Saingamsook; Thanari Phanitchakun; Ralph E Harbach; Pradya Somboon

doi:10.1093/jme/tjad089

A multiplex PCR assay for the identification of 5 sibling species of the Lindesayi Complex of the genus Anopheles (Diptera: Culicidae) in Bhutan

J Med Entomol. 2023 Sep 12;60(5):924-930. doi: 10.1093/jme/tjad089.

Authors

Li Zhou¹, Rinsin Namgay², Atiporn Saeung¹, Jassada Saingamsook¹, Thanari Phanitchakun¹, Ralph E Harbach³, Pradya Somboon¹

Affiliations

¹ Center of Insect Vector Study, Department of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
² Vector-borne Disease Control Programme, Ministry of Health, Gelephu 31101, Bhutan.
³ Department of Science, Natural History Museum, Cromwell Road, London SW7 5BD, UK.

PMID: 37417304
DOI: 10.1093/jme/tjad089

Abstract

Five sibling species of the Lindesayi Complex of the genus Anopheles have been discovered in Bhutan: An. druki Somboon, Namgay & Harbach, An. himalayensis Somboon, Namgay & Harbach, An. lindesayi Giles, An. lindesayi species B, and An. thimphuensis Somboon, Namgay & Harbach. The species are morphologically similar in the adult and/or immature stages. This study aimed to develop a multiplex PCR assay to identify the 5 species. Allele-specific primers were designed for specific nucleotide segments of ITS2 sequences previously reported for each species. The assay provided products of 183 bp for An. druki, 338 bp for An. himalayensis, 126 bp for An. lindesayi, 290 bp for An. lindesayi species B, and 370 bp for An. thimphuensis. The use of the assay produced consistent results. The assay is relatively inexpensive, enables the rapid identification of a large number of specimens, and will foster further studies of the Lindesayi Complex.

Keywords: ITS2; PCR product; molecular method; mosquito; primer design.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Anopheles*
Bhutan
Multiplex Polymerase Chain Reaction