A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group.

Koekemoer, Lizette L.; Kamau, Luna; Hunt, Richard H.; Coetzee, Maureen

doi:10.4269/ajtmh.2002.66.804

Cited by 501 publications

(504 citation statements)

References 15 publications

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“…Two seasonal collections were needed to generate adequate numbers to test the full spectrum of insecticides. Eggs were collected, and female F0 mosquitoes used for oviposition were species-identified using funestus complex mixture PCR (27). Insecticide resistance was assessed using 2-to 5-d-old F1 adult mosquitoes.…”

Section: Methodsmentioning

confidence: 99%

Restriction to gene flow is associated with changes in the molecular basis of pyrethroid resistance in the malaria vector Anopheles funestus

Barnes

Irving

Chiumia

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Resistance to pyrethroids, the sole insecticide class recommended for treating bed nets, threatens the control of major malaria vectors, including Anopheles funestus. Effective management of resistance requires an understanding of the dynamics and mechanisms driving resistance. Here, using genome-wide transcription and genetic diversity analyses, we show that a shift in the molecular basis of pyrethroid resistance in southern African populations of this species is associated with a restricted gene flow. Across the most highly endemic and densely populated regions in Malawi, An. funestus is resistant to pyrethroids, carbamates, and organochlorides. Genome-wide microarray-based transcription analysis identified overexpression of cytochrome P450 genes as the main mechanism driving this resistance. The most up-regulated genes include cytochrome P450s (CYP) CYP6P9a, CYP6P9b and CYP6M7. However, a significant shift in the overexpression profile of these genes was detected across a south/north transect, with CYP6P9a and CYP6P9b more highly overexpressed in the southern resistance front and CYP6M7 predominant in the northern front. A genome-wide genetic structure analysis of southern African populations of An. funestus from Zambia, Malawi, and Mozambique revealed a restriction of gene flow between populations, in line with the geographical variation observed in the transcriptomic analysis. Genetic polymorphism analysis of the three key resistance genes, CYP6P9a, CYP6P9b, and CYP6M7, support barriers to gene flow that are shaping the underlying molecular basis of pyrethroid resistance across southern Africa. This barrier to gene flow is likely to impact the design and implementation of resistance management strategies in the region.

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Section: Methodsmentioning

confidence: 99%

Restriction to gene flow is associated with changes in the molecular basis of pyrethroid resistance in the malaria vector Anopheles funestus

Barnes

Irving

Chiumia

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…Samples were transported to the Montpellier laboratory and stored at 4°C. Infected females belonging to the A. gambiae complex and A. funestus group were identified to species by PCR diagnostic assays (44,45).…”

Section: Methodsmentioning

confidence: 99%

Population genetic structure of Plasmodium falciparum in the two main African vectors, Anopheles gambiae and Anopheles funestus

Annan

Durand

Ayala

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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We investigated patterns of genetic diversity of Plasmodium falciparum associated with its two main African vectors: Anopheles gambiae and Anopheles funestus. We dissected 10,296 wildcaught mosquitoes from three tropical sites, two in Cameroon (Simbock and Tibati, separated by 320 km) and one in Kenya (Rota, >2,000 km from the other two sites). We assayed seven microsatellite loci in 746 oocysts from 183 infected mosquito guts. Genetic polymorphism was very high in parasites isolated from both vector species. The expected heterozygosity (HE) was 0.79 in both species; the observed heterozygosities (HO) were 0.32 in A. funestus and 0.42 in A. gambiae, indicating considerable inbreeding within both vector species. Mean selfing (s) between genetically identical gametes was s ‫؍‬ 0.33. Differences in the rate of inbreeding were statistically insignificant among sites and between the two vector species. As expected, because of the high rate of inbreeding, linkage disequilibrium was very high; it was significant for all 21 loci pairs in A. gambiae and for 15 of 21 pairs in A. funestus, although only two pairwise comparisons were between loci on the same chromosome. Overall, the genetic population structure of P. falciparum, as evaluated by F statistics, was predominantly clonal rather than panmictic, a population structure that facilitates the spread of antimalarial drug and vaccine resistance and thus may impair the effectiveness of malaria control efforts. malaria ͉ epidemiology ͉ evolutionary genetics ͉ Cameroon ͉ Kenya M alaria is the most significant and widespread vectortransmitted human disease, accounting yearly for several hundred million clinical cases and Ͼ2 million deaths, mostly affecting young children and pregnant women in subSaharan Africa (1). Prevention and cure of the disease are major publichealth challenges that are tackled by using several strategies, among them vector control. The transmission of Plasmodium falciparum, the agent of malignant malaria, involves a complex vectorial system consisting of Ϸ10 Anopheles species, colonizing different ecoclimatic settings, regions, and seasons in strongly variable relative abundances (2-5).The two most important vectors of malignant malaria in Africa are Anopheles gambiae and Anopheles funestus because of their widespread distribution, highly anthropophilic and endophilic behavior, and long life spans (6, 7). A. gambiae is the most important vector throughout Africa and the most extensively studied Anopheles species (8). The effectiveness of malaria transmission emerges from the complementary ecoclimatic attributes and seasonal patterns of both species. A. funestus breeds in permanent larval sites that enable this species, in regions of seasonal transmission, to extend parasite transmission far into the dry season, after the temporary breeding pools of A. gambiae have dried out (7, 9-11).Differences in the biology and ecology of these two main vectors might entail a differential impact on the genetic composition of the P. falciparum populations they harbo...

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“…Prior to analysis by PCR, genomic DNA was diluted 1:10 in H 2 O ($5 ng/ml). Identification of A. funestus sensu stricto apart from closely related species with similar or identical adult morphology was achieved by an rDNA-based PCR assay (Koekemoer et al 2002) as modified by Michel et al (2005a). Sporozoite infection was assayed by nested PCR targeting the MspI gene (Ranford-Cartwright et al 1993;Michel et al 2005a).…”

Section: Methodsmentioning

confidence: 99%

Divergence With Gene Flow in Anopheles funestus From the Sudan Savanna of Burkina Faso, West Africa

et al. 2006

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Anopheles funestus is a major vector of malaria across Africa. Understanding its complex and nonequilibrium population genetic structure is an important challenge that must be overcome before vector populations can be successfully perturbed for malaria control. Here we examine the role of chromosomal inversions in structuring genetic variation and facilitating divergence in Burkina Faso, West Africa, where two incipient species (chromosomal forms) of A. funestus, defined principally by rearrangements of chromosome 3R, have been hypothesized. Sampling across an $300-km east-west transect largely contained within the Sudan-Savanna ecoclimatic zone, we analyzed chromosomal inversions, 16 microsatellite loci distributed genomewide, and 834 bp of the mtDNA ND5 gene. Both molecular markers revealed high genetic diversity, nearly all of which was accounted for by within-population differences among individuals, owing to recent population expansion. Across the study area there was no correlation between genetic and geographic distance. Significant genetic differentiation found between chromosomal forms on the basis of microsatellites was not genomewide but could be explained by chromosome 3R alone on the basis of loci inside and near inversions. These data are not compatible with complete reproductive isolation but are consistent with differential introgression and sympatric divergence between the chromosomal forms, facilitated by chromosome 3R inversions.

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A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group.

Cited by 501 publications

References 15 publications

Restriction to gene flow is associated with changes in the molecular basis of pyrethroid resistance in the malaria vector Anopheles funestus

Restriction to gene flow is associated with changes in the molecular basis of pyrethroid resistance in the malaria vector Anopheles funestus

Population genetic structure of Plasmodium falciparum in the two main African vectors, Anopheles gambiae and Anopheles funestus

Divergence With Gene Flow in Anopheles funestus From the Sudan Savanna of Burkina Faso, West Africa

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