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PCR-RFLP

A PCR-RFLP method for genotyping of inversion 2Rc in Anopheles coluzzii

March 23, 2021 - 14:42 -- Open Access
Author(s): 
Montanez-Gonzalez R, Vallera AC, Besansky NJ, et al.
Reference: 
Parasit Vectors. 2021 Mar 22;14(1):174

Genotyping of polymorphic chromosomal inversions in malaria vectors such as An. coluzzii Coetzee & Wilkerson is important, both because they cause cryptic population structure that can mislead vector analysis and control and because they influence epidemiologically relevant eco-phenotypes. The conventional cytogenetic method of genotyping is an impediment because it is labor intensive, requires specialized training, and can be applied only to one gender and developmental stage. Here, we circumvent these limitations by developing a simple and rapid molecular method of genotyping inversion 2Rc in An. coluzzii that is both economical and field-friendly. This inversion is strongly implicated in temporal and spatial adaptations to climatic and ecological variation, particularly aridity.

Highly specific PCR-RFLP assays for karyotyping the widespread 2Rb inversion in malaria vectors of the Anopheles gambiae complex

January 14, 2020 - 10:08 -- Open Access
Author(s): 
Montanez-Gonzalez R, Pichler V, Calzetta M, Love RR, Vallera A, Schaecher L, Caputo B, Pombi M, Petrarca V, Della Torre A, Besansky NJ
Reference: 
Parasites & Vectors, Volume 13, 16 (2020)

Chromosomal inversion polymorphisms play a role in adaptation to heterogeneous environments. Inversion polymorphisms are implicated in the very high ecological flexibility of the three main malaria vector species of the Afrotropical Anopheles gambiae complex, facilitating the exploitation of anthropogenic environmental modifications and promoting a strong association with humans. In addition to extending the species’ spatial and temporal distribution, inversions are associated with epidemiologically relevant mosquito behavior and physiology, underscoring their medical importance. We here present novel PCR-RFLP based assays strongly predictive of genotype for the cosmopolitan 2Rb inversion in An. coluzzii and An. gambiae, a development which overcomes the numerous constraints inherent to traditional cytological karyotyping.

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