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A panel of ten microsatellite loci for the Chagas disease vector Rhodnius prolixus (Hemiptera: Reduviidae).

Fitzpatrick S, Watts PC, Feliciangeli MD, Miles MA, Kemp SJ - Infect. Genet. Evol. (2008)

Bottom Line: Vector control failure in Venezuela may be due to the invasion of houses by silvatic populations of R. prolixus found in palms.As part of a study to determine if domestic and silvatic populations of R. prolixus are isolated, thus clarifying the role of silvatic populations in maintaining house infestations, we constructed three partial genomic microsatellite libraries.Allele numbers per locus ranged from three to twelve, with observed and expected heterozygosity ranging from 0.26 to 0.55 and 0.32 to 0.66.

View Article: PubMed Central - PubMed

Affiliation: Pathogen and Molecular Biology Unit, Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E7HT, UK. sinead.fitzpatrick@lshtm.ac.uk

ABSTRACT
Rhodnius prolixus is the main vector of Chagas disease in Venezuela, where it is found colonising rural housing consisting of unplastered adobe walls with palm and/or metal roofs. Vector control failure in Venezuela may be due to the invasion of houses by silvatic populations of R. prolixus found in palms. As part of a study to determine if domestic and silvatic populations of R. prolixus are isolated, thus clarifying the role of silvatic populations in maintaining house infestations, we constructed three partial genomic microsatellite libraries. A panel of ten dinucleotide polymorphic microsatellite markers was selected for genotyping. Allele numbers per locus ranged from three to twelve, with observed and expected heterozygosity ranging from 0.26 to 0.55 and 0.32 to 0.66. The microsatellite markers presented here will contribute to the control of Chagas disease in Venezuela and Colombia through the provision of population information that may allow the design of improved control strategies.

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The distribution of alleles at each tested locus.
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fig1: The distribution of alleles at each tested locus.

Mentions: A total of 555 R. prolixus specimens collect from silvatic, domestic and peridomestic sites in five Venezuelan States were analysed using this panel of ten microsatellites. Results of the population analysis are reported elsewhere (Fitzpatrick et al., 2008). The number of alleles per locus ranged from 3 (LIST14-042) up to 12 (LIST14-010) (Table 1, Fig. 1). In this study 54 specimens failed to amplify at a single locus, when all other loci for these specimens generated product. Two specimens failed at two loci. All 10 loci experienced such non amplification, with greatest numbers occurring at LIST14-010, LIST14-037, LIST14-013 (11 and 10 specimens, respectively). Null alleles are only obvious in homozygotes where there is a failure to amplify product. Heterozygotes with a single allelle could be mistakenly identified as homozygotes. Observed and expected heterozygosity varied between 0.26–0.55 and 0.32–0.66 respectively (Table 1). Seven loci showed significant deviations from expected Hardy-Weinberg conditions after sequential Bonferroni due to heterozygosity deficiency. However this might be expected as specimens were pooled from 33 populations from 5 States in Venezuela. The number of polymorphic loci in populations ranged from 6 to 10, with 85% of all populations polymorphic at all loci. Fisher's exact test across each pair of loci in the pooled populations detected significant linkage at two locus pairs, LIST14-056 with LIST14-013 and LIST14-056 with LIST14-076, both remained significant after Bonferroni correction. Significant linkage disequilibrium was detected in 96 locus pairs in 30 populations, however only 18 pairs remained significant after sequential Bonferroni correction: LIST14-017 with LIST14-042 (1 population), LIST14-010 with LIST14-013 (1 population), LIST14-010 and LIST14-025 (1 population) and LIST14-056 with LIST14-076 (15 populations). Due to the consistent linkage pattern exhibited by LIST14-056 with LIST14-076, locus LIST14-076 was excluded from further population analysis (Fitzpatrick et al., 2008). Cluster analysis of all 10 loci did not detect shared flanking regions (BioEdit, Hall 1999).


A panel of ten microsatellite loci for the Chagas disease vector Rhodnius prolixus (Hemiptera: Reduviidae).

Fitzpatrick S, Watts PC, Feliciangeli MD, Miles MA, Kemp SJ - Infect. Genet. Evol. (2008)

The distribution of alleles at each tested locus.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651431&req=5

fig1: The distribution of alleles at each tested locus.
Mentions: A total of 555 R. prolixus specimens collect from silvatic, domestic and peridomestic sites in five Venezuelan States were analysed using this panel of ten microsatellites. Results of the population analysis are reported elsewhere (Fitzpatrick et al., 2008). The number of alleles per locus ranged from 3 (LIST14-042) up to 12 (LIST14-010) (Table 1, Fig. 1). In this study 54 specimens failed to amplify at a single locus, when all other loci for these specimens generated product. Two specimens failed at two loci. All 10 loci experienced such non amplification, with greatest numbers occurring at LIST14-010, LIST14-037, LIST14-013 (11 and 10 specimens, respectively). Null alleles are only obvious in homozygotes where there is a failure to amplify product. Heterozygotes with a single allelle could be mistakenly identified as homozygotes. Observed and expected heterozygosity varied between 0.26–0.55 and 0.32–0.66 respectively (Table 1). Seven loci showed significant deviations from expected Hardy-Weinberg conditions after sequential Bonferroni due to heterozygosity deficiency. However this might be expected as specimens were pooled from 33 populations from 5 States in Venezuela. The number of polymorphic loci in populations ranged from 6 to 10, with 85% of all populations polymorphic at all loci. Fisher's exact test across each pair of loci in the pooled populations detected significant linkage at two locus pairs, LIST14-056 with LIST14-013 and LIST14-056 with LIST14-076, both remained significant after Bonferroni correction. Significant linkage disequilibrium was detected in 96 locus pairs in 30 populations, however only 18 pairs remained significant after sequential Bonferroni correction: LIST14-017 with LIST14-042 (1 population), LIST14-010 with LIST14-013 (1 population), LIST14-010 and LIST14-025 (1 population) and LIST14-056 with LIST14-076 (15 populations). Due to the consistent linkage pattern exhibited by LIST14-056 with LIST14-076, locus LIST14-076 was excluded from further population analysis (Fitzpatrick et al., 2008). Cluster analysis of all 10 loci did not detect shared flanking regions (BioEdit, Hall 1999).

Bottom Line: Vector control failure in Venezuela may be due to the invasion of houses by silvatic populations of R. prolixus found in palms.As part of a study to determine if domestic and silvatic populations of R. prolixus are isolated, thus clarifying the role of silvatic populations in maintaining house infestations, we constructed three partial genomic microsatellite libraries.Allele numbers per locus ranged from three to twelve, with observed and expected heterozygosity ranging from 0.26 to 0.55 and 0.32 to 0.66.

View Article: PubMed Central - PubMed

Affiliation: Pathogen and Molecular Biology Unit, Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E7HT, UK. sinead.fitzpatrick@lshtm.ac.uk

ABSTRACT
Rhodnius prolixus is the main vector of Chagas disease in Venezuela, where it is found colonising rural housing consisting of unplastered adobe walls with palm and/or metal roofs. Vector control failure in Venezuela may be due to the invasion of houses by silvatic populations of R. prolixus found in palms. As part of a study to determine if domestic and silvatic populations of R. prolixus are isolated, thus clarifying the role of silvatic populations in maintaining house infestations, we constructed three partial genomic microsatellite libraries. A panel of ten dinucleotide polymorphic microsatellite markers was selected for genotyping. Allele numbers per locus ranged from three to twelve, with observed and expected heterozygosity ranging from 0.26 to 0.55 and 0.32 to 0.66. The microsatellite markers presented here will contribute to the control of Chagas disease in Venezuela and Colombia through the provision of population information that may allow the design of improved control strategies.

Show MeSH
Related in: MedlinePlus