A NEW HIGH-PERFORMANCE PCR DIAGNOSTIC FOR THE DETECTION OF PYRETHROID KNOCKDOWN RESISTANCE KDR IN ANOPHELES GAMBIAE

FREDERIC TRIPET Department of Entomology, University of California Davis, Davis, California

Search for other papers by FREDERIC TRIPET in
Current site
Google Scholar
PubMed
Close
,
JENNIFER WRIGHT Department of Entomology, University of California Davis, Davis, California

Search for other papers by JENNIFER WRIGHT in
Current site
Google Scholar
PubMed
Close
, and
GREG LANZARO Department of Entomology, University of California Davis, Davis, California

Search for other papers by GREG LANZARO in
Current site
Google Scholar
PubMed
Close
Restricted access

Monitoring the spread of the knockdown resistance allele kdr in areas of extensive pyrethroid use is critical to vector-control projects. Currently available methods for detecting kdr from DNA samples are characterized by poor amplification, time-consuming steps, and primers that exhibit frequent null alleles. We describe a new PCR diagnostic that uses fluorescent primers based on conserved priming sites and enables simple detection of the kdr allele on a sequencer. Using samples from a West African Anopheles gambiae population, we show that the new PCR yielded significantly higher rates of amplification and more accurate estimates of kdr frequency. The method works equally well for the leucine to phenylalanine substitution found in West Africa and the East African leucine to serine substitution.

Author Notes

Reprint requests: Frederic Tripet, Center for Applied Entomology and Parasitology, School of Life Sciences, Keele University, Stafford-shire, ST5 5BG, UK, E-mail: frtripet@yahoo.com.
  • 1

    World Health Organization, 1999. The World Health Report 1999: Making a difference. Geneva, World Health Organization.

    • PubMed
    • Export Citation
  • 2

    Marshall E, 2001. A renewed assault on an old and deadly foe. Science 290 :428–430.

  • 3

    Coluzzi M, Sabatini A, 1967. Cytogenetic observations on species A and B of the Anopheles gambiae complex. Parassitologia 9 :71–88.

  • 4

    Davidson G, Paterson HE, Coluzzi M, Mason GF, Micks DW, 1967. The Anopheles gambiae complex. Wright JW, Pal R, eds. Genetics of Insect Vectors of Disease. Amsterdam: Elsevier, 211–250.

    • PubMed
    • Export Citation
  • 5

    Coetzee M, Craig M, Le Sueur D, 2000. Distribution of African malaria mosquitoes belonging to the Anopheles gambiae complex. Parasitol Today 16 :74–77.

  • 6

    Bryan JH, Di Deco MA, Petrarca V, Coluzzi M, 1982. Inversion polymorphism and incipient speciation in Anopheles gambiae s. str. in the Gambiae, West Africa. Genetica 59 :167–176.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7

    Touré YT, Petrarca V, Coluzzi M, 1983. Nueva entita del complesso Anopheles gambiae in Mali. Parrasitologia 25 :367–370.

  • 8

    Coluzzi M, Petrarca V, Di Deco MA, 1985. Chromosomal inversion intergradation and incipient speciation in Anopheles gambiae. Bollettino di Zoologia 52 :45–63.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9

    Favia G, Della Torre A, Bagyaoko M, Lanfrancotti A, Sagnon NF, Toure YT, Coluzzi M, 1997. Molecular identifications of sympatric chromosomal forms of Anopheles gambiae and further evidence of their reproductive isolation. Insect Mol Biol 6 :377–383.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 10

    Favia G, Lanfrancotti A, Spanos L, Sidén-Kiamos I, Louis C, 2001. Molecular characterization of ribosomal DNA (rDNA) polymorphisms discriminating among chromosomal forms of Anopheles gambiae s.s. Insect Mol Biol 10 :19–23.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 11

    Della Torre A, Costantini C, Besansky NJ, Caccone A, Petrarca V, Powell JR, Coluzzi M, 2002. Speciation within Anopheles gambiae - the glass is half full. Science 298 :115–117.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12

    Gentile G, Della Torre A, Maegga B, Powell JR, Caccone A, 2002. Genetic differentiation in the African malaria vector, Anopheles gambiae s.s., and the problem of taxonomic status. Genetics 161 :1561–1578.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13

    Wondji C, Simard F, Fontenille D, 2002. Evidence for genetic differentiation between the molecular forms M and S within the Forest chromosomal form of Anopheles gambiae in an area of sympatry. Insect Mol Biol 11 :11–19.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14

    Choi HW, Breman JG, Teutsch SM, Liu S, Hightower AW, Sexton JD, 1995. The effectiveness of insecticide-impregnated bed nets in reducing cases of malaria infection: a meta-analysis of published results. Am J Trop Med Hyg 52 :377–382.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15

    Mittal PK, Adak T, Sharma VP, 1991. Acute toxicity of certain organochlorine, organophosphorus, synthetic pyrethroid and microbial insecticides to the mosquito fish Gambusia affinis (Baird and Girard). Indian J Malariol 28 :167–170.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16

    Hemingway J, Ranson H, 2000. Insecticide resistance in insect vectors of human disease. Annu Rev Entomol 45 :371–391.

  • 17

    Ranson H, Jensen B, Vulule JM, Wang X, Hemingway J, Collins FH, 2000. Identification of a point mutation in the voltage-gated sodium channel gene of Kenyan Anopheles gambiae associated with resistance to DDT and pyrethroids. Insect Mol Biol 9 :491–497.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 18

    Martinez-Torres D, Chandre F, Williamson MS, Darriet F, Bergé JB, Devonshire AL, Guillet P, Pasteur N, Pauron D, 1998. Molecular characterization of pyrethroid knockdown resistance (kdr) in the major malaria vector Anopheles gambiae s.s. Insect Mol Biol 7 :179–184.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19

    Elissa N, Mouchet J, Riviere F, Meunier JY, Yao K, 1993. Resistance of Anopheles gambiae s.s. to pyrethroids in Cote d’Ivoire. Ann Soc Belg Med Trop 73 :291–294.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20

    Weill M, Chandre F, Brengues C, Manguin S, Akogbeto M, Pasteur N, Guillet P, Raymond M, 2000. The kdr mutation occurs in the Mopti form of Anopheles gambiae s.s. through introgression. Insect Mol Biol 9 :451–455.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21

    Diabate A, Baldet T, Chandre C, Dabire KR, Kengne P, Guiguemde TR, Simard F, Guillet P, Hemingway J, Hougard JM, 2003. Kdr mutation, a genetic marker to assess events of introgression between the molecular M and S forms of Anopheles gambiae (Diptera: Culicidae) in the tropical savannah area of West Africa. J Med Entomol 40 :195–198.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22

    Diabate A, Brengues C, Baldet T, Dabire KR, Hougard JM, Akogbeto M, Kengne P, Simard F, Guillet P, Hemingway J, Chandre F, 2004. The spread of the Leu-Phe kdr mutation through Anopheles gambiae complex in Burkina Faso: genetic introgression and de novo phenomena. Trop Med Int Health 9 :1267–1273.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23

    Gentile G, Santolamazza F, Fanello C, Petrarca V, Caccone A, della Torre A, 2004. Variation in an intron sequence of the voltage-gated sodium channel gene correlates with genetic differentiation between Anopheles gambiae s.s. molecular forms. Insect Mol Biol 13 :371–377.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24

    Fanello C, Petrarca V, della Torre A, Santolamazza F, Dolo G, Coulibaly M, Alloueche A, Curtis CF, Touré YT, Coluzzi M, 2003. The pyrethroid knock-down resistance gene in the Anopheles gambiae complex in Mali and further indication of incipient speciation within An. gambiae s.s. Insect Mol Biol 12 :241–245.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25

    Yawson AE, McCall PJ, Wilson MD, Donnely MJ, 2004. Species abundance and insecticide resistance of Anopheles gambiae in selected areas of Ghana and Burkina Faso. Med. Vet. Entomol 18 :372–377.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 26

    Stump AD, Atieli FK, Vulule JM, Besansky NJ, 2004. Dynamics of the pyrethroid knockdown resistance allele in western Kenyan populations of Anopheles gambiae in response to insecticide-treated bed net trials. Am J Trop Med Hyg 70 :591–596.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 27

    Lanzaro GC, Tripet F, 2003. Gene flow among populations of Anopheles gambiae: a critical review. Takken W, Scott TW, eds. Ecological Aspects for the Application of Genetically Modified Mosquitoes. Wageningen, The Netherlands: Frontis Press, 109–132.

    • PubMed
    • Export Citation
  • 28

    Reimer LJ, Tripet F, Slotman M, Spielman A, Fondjo E, Lanzaro GC, 2005. An unusual distribution of the kdr gene among populations of Anopheles gambiae on the island of Bioko, Equatorial Guinea. Insect Mol Biol 14 :683–688.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 29

    Kolaczinski JH, Fanello C, Hervé J-P, Conway DJ, Carneval P, Curtis F, 2000. Experimental and molecular genetic analysis of the impact of pyrethroid and non-pyrethroid insecticide impregnated bednets for mosquito control in an area of pyrethroid resistance. Bull Entomol Res 9 :125–132.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 30

    Lynd A, Ranson H, McCall PJ, Randle NP, Black WC IV, Walker ED, Donelly MJ, 2005. A simplified high-throughput method for pyrethroid knock-down resistance (kdr) detection in Anopheles gambiae. Malaria J 4 :16.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31

    Scott JA, Brogdon WG, Collins FH, 1993. Identification of single specimens of the Anopheles gambiae complex by the polymerase chain reaction. Am J Trop Med Hyg 49 :520–529.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 32

    Fanello C, Santolamazza F, Della Torre A, 2002. Simultaneous identification of species and molecular forms of the Anopheles gambiae complex by PCR-RFLP. Med Vet Entomol 16 :461–464.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 33

    Rychlik W, 2002. Oligo—Primer Analysis software. Version 6. Molecular Insights, West Cascade, CO.

    • PubMed
    • Export Citation
Past two years Past Year Past 30 Days
Abstract Views 54 31 2
Full Text Views 89 3 1
PDF Downloads 40 3 1
 
 
 
 
Affiliate Membership Banner
 
 
Research for Health Information Banner
 
 
CLOCKSS
 
 
 
Society Publishers Coalition Banner
Save