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Detection and monitoring of insecticide resistance mutations in Anopheles gambiae: individual vs. pooled specimens

Mavridis, Konstantinos and Wipf, Nadja and Müller, Pie and Traoré, Mohamed M. and Muller, Gunter and Vontas, John. (2018) Detection and monitoring of insecticide resistance mutations in Anopheles gambiae: individual vs. pooled specimens. Genes, 9 (10). E479.

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Official URL: https://edoc.unibas.ch/65728/

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Abstract

Bioassays and molecular diagnostics are routinely used for the monitoring of malaria vector populations to support insecticide resistance management (IRM), guiding operational decisions on which insecticides ought to be used for effective vector control. Previously developed TaqMan assays were optimised to distinguish the wild-type L1014 from the knockdown resistance (; kdr; ) point mutations 1014F and 1014S (triplex reaction), and the N1575 wild-type from the point mutation 1575Y (duplex reaction). Subsequently, artificial pools of; Anopheles gambiae (An. gambiae); specimens with known genotypes of L1014F, L1014S, and N1575Y were created, nucleic acids were extracted, and; kdr; mutations were detected. These data were then used to define a linear regression model that predicts the allelic frequency within a pool of mosquitoes as a function of the measured ΔCt values (Ct mutant - Ct wild type probe). Polynomial regression models showed; r; 2; values of >0.99 (; p; < 0.05). The method was validated with populations of variable allelic frequencies, and found to be precise (1.66⁻2.99%), accurate (3.3⁻5.9%), and able to detect a single heterozygous mosquito mixed with 9 wild type individuals in a pool of 10. Its pilot application in field-caught samples showed minimal differences from individual genotyping (0.36⁻4.0%). It allowed the first detection of the super-; kdr; mutation N1575Y in; An. gambiae; from Mali. Using pools instead of individuals allows for more efficient resistance allele screening, facilitating IRM.
Faculties and Departments:09 Associated Institutions > Swiss Tropical and Public Health Institute (Swiss TPH)
09 Associated Institutions > Swiss Tropical and Public Health Institute (Swiss TPH) > Department of Epidemiology and Public Health (EPH) > Health Interventions > Vector Control Centre (Müller)
UniBasel Contributors:Müller, Pie and Wipf, Nadja
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Multidisciplinary Digital Publishing Institute
ISSN:2073-4425
Note:Publication type according to Uni Basel Research Database: Journal article
Language:English
Identification Number:
Last Modified:17 Oct 2018 13:18
Deposited On:17 Oct 2018 13:18

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