Author:
- Introduction
- Materials and Methods
- Results
- kdr-associated mutations.
- VGSC coding sequence analysis.
- Repeated synonymous SNPs.
- Repeated nonsynonymous SNPs.
- VGSC sequence comparisons outside the An. punctulatus species group.
- Test for evidence of selection.
- VGSC intron sequence analysis.
- Phylogenetic comparison of An. punctulatus sibling species using VGSC sequence.
- Post-PCR LDR-FMA design and validation.
- Application of VGSC multiplex assay: analysis of mosquito collections for kdr mutation and species determination.
- Discussion
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Multiplex Assay for Species Identification and Monitoring of Insecticide Resistance in Anopheles punctulatus Group Populations of Papua New Guinea
Cara N. Henry-Halldin
Cara N. Henry-Halldin
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Kogulan Nadesakumaran
Kogulan Nadesakumaran
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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John Bosco Keven
John Bosco Keven
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Allison M. Zimmerman
Allison M. Zimmerman
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Peter Siba
Peter Siba
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Ivo Mueller
Ivo Mueller
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Manuel W. Hetzel
Manuel W. Hetzel
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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James W. Kazura
James W. Kazura
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Edward Thomsen
Edward Thomsen
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Lisa J. Reimer
Lisa J. Reimer
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Peter A. Zimmerman
Peter A. Zimmerman
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio; Papua New Guinea Institute of Medical Research, Madang, MADANG, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, EASTERN HIGHLANDS, Papua New Guinea; School of Population Health, University of Queensland, Brisbane, Australia
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Anopheles punctulatus sibling species (An. punctulatus s.s., Anopheles koliensis, and Anopheles farauti species complex [eight cryptic species]) are principal vectors of malaria and filariasis in the Southwest Pacific. Given significant effort to reduce malaria and filariasis transmission through insecticide-treated net distribution in the region, effective strategies to monitor evolution of insecticide resistance among An. punctulatus sibling species is essential. Mutations in the voltage-gated sodium channel (VGSC) gene have been associated with knock-down resistance (kdr) to pyrethroids and DDT in malarious regions. By examining VGSC sequence polymorphism we developed a multiplex assay to differentiate wild-type versus kdr alleles and query intron-based polymorphisms that enable simultaneous species identification. A survey including mosquitoes from seven Papua New Guinea Provinces detected no kdr alleles in any An. punctulatus species. Absence of VGSC sequence introgression between species and evidence of geographic separation within species suggests that kdr must be monitored in each An. punctulatus species independently.
Author Notes
Financial support: This study was supported by grants from National Institutes of Health (AI065717) and Fogarty International Center (TW007872, TW007377, and TW007735). Mosquito collection was also supported by the Global Fund to Fight Aids, Tuberculosis and Malaria Round 3 malaria grant to PNG.
Authors' addresses: Cara N. Henry-Halldin, Kogulan Nadesakumaran, Allison M. Zimmerman, James W. Kazura, and Peter A. Zimmerman, Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, OH, E-mails: vgx5@cdc.gov, kxn80@case.edu, amz23@case.edu, jxk14@case.edu, and paz@case.edu. John Bosco Keven, Edward Thomsen, and Lisa J. Reimer, Papua New Guinea Institute of Medical Research-Madang, Madang, Papua New Guinea, E-mails: jbkeven@gmail.com, edward.thomsen@case.edu, and lisa.reimer@case.edu. Peter Siba, Ivo Mueller, and Manuel W. Hetzel, Papua New Guinea Institute of Medical Research-Goroka, Eastern Highlands, Papua New Guinea, E-mails: peter.siba@pngimr.org.pg, ivomueller@fastmail.fm, and manuel.hetzel@pngimr.org.pg.
- Introduction
- Materials and Methods
- Results
- kdr-associated mutations.
- VGSC coding sequence analysis.
- Repeated synonymous SNPs.
- Repeated nonsynonymous SNPs.
- VGSC sequence comparisons outside the An. punctulatus species group.
- Test for evidence of selection.
- VGSC intron sequence analysis.
- Phylogenetic comparison of An. punctulatus sibling species using VGSC sequence.
- Post-PCR LDR-FMA design and validation.
- Application of VGSC multiplex assay: analysis of mosquito collections for kdr mutation and species determination.
- Discussion
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