• 1

    Coluzzi M, Sabatini A, Petrarca V, di Deco MA, 1979. Chromosomal differentiation and adaptation to human environment in the Anopheles gambiae complex. Trans R Soc Trop Med Hyg 73 :483–497.

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  • 2

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

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  • 3

    Coluzzi M, Sabatini A, della Torre A, Di Deco MA, Petrarca V, 2002. The polytene chromosome map of Anopheles gambiae: cytotaxonomic findings, patterns of rearrangements, evolutionary and malariological implications. Science 298 :1415–1418.

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  • 4

    Touré YT, Petrarca V, Traoré SF, Coulibaly A, Maiga HM, Sankaré O, Sow M, Di Deco MA, Coluzzi M, 1994. Ecological genetic studies in the chromosomal form Mopti of Anopheles gambiae s.s. in Mali, west Africa. Genetica 94 :213–223.

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  • 5

    Touré YT, Petrarca V, Traoré SF, Coulibaly A, Maiga HM, Sankaré O, Sow M, Di Deco MA, Coluzzi M, 1998. Distribution and inversion polymorphism of chromosomally recognized taxa of the Anopheles gambiae complex in Mali, west Africa. Parassitologia 40 :477–511.

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  • 6

    Powell JR, Petrarca V, della Torre A, Caccone A, Coluzzi M, 1999. Population structure, speciation, and introgression in the Anopheles gambiae complex.Parassitologia 41 :101–114.

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  • 7

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

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  • 8

    Mukabayire O, Touré YT, Coluzzi M, Besansky NJ, 2001. Patterns of DNA sequence variation in chromosomally recognized taxa of Anopheles gambiae: evidence from rDNA, white and tryptophan oxygenase genes. Insect Mol Biol 10 :33–46.

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  • 9

    della Torre A, Fanello C, Akogbeto M, Dossouvovo J, Favia G, Petrarca V, Coluzzi. M, 2001. Cytogenetic and molecular evidence of incipient speciation within Anopheles gambiae s.s. in west Africa. Insect Mol Biol 10 :9–18.

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  • 10

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

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  • 11

    Favia G, Dimopoulos G, della Torre A, Touré YT, Coluzzi M, Louis C, 1994. Polymorphisms detected by random PCR distinguishes between different chromosomal forms of Anopheles gambiae.Proc Natl Acad Sci USA 91 :10315–10319.

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  • 12

    Favia G, della Torre A, Bagayoko M, Lanfrancotti A, Sagnon N’F, Touré YT, Coluzzi M, 1997. Molecular identification of sympatric chromosomal forms of Anopheles gambiae and further evidence of their reproductive isolation. Insect Mol Biol 6 :377–383.

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  • 13

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

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    Gentile G, Slotman M, Ketmaier V, Powell JR, Caccone A, 2001. Attempts to molecularly distinguish cryptic taxa in Anopheles gambiae s.s. Insect Mol Biol 10 :25–32.

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    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.

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  • 16

    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.

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  • 17

    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 : (in press).

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  • 18

    Edillo FE, Touré YT, Lanzaro GC, Dolo G, Taylor CE, 2002. Spatial and habitat distribution of Anopheles gambiae and Anopheles arabiensis (Diptera:Culicidae) in Banambani village, Mali. J Med Entomol 39 :70–77.

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  • 19

    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.

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  • 20

    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 :461–466.

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  • 21

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

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SHORT REPORT: A NEW POLYMERASE CHAIN REACTION-RESTRICTION FRAGMENT LENGTH POLYMORPHISM METHOD TO IDENTIFY ANOPHELES ARABIENSIS FROM AN. GAMBIAE AND ITS TWO MOLECULAR FORMS FROM DEGRADED DNA TEMPLATES OR MUSEUM SAMPLES

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  • 1 Department of Ecology and Evolutionary Biology and Yale Institute for Biospheric Studies, Yale University, New Haven, Connecticut; Sezione di Parassitologia, Dipartimento di Scienze di Sanità Pubblica, Università di Roma La Sapienza, Rome, Italy

We present a polymerase chain reaction-restriction fragment length polymorphism method to simultaneously distinguish the two Anopheles gambiae M and S molecular forms and Anopheles arabiensis. This method uses different diagnostic sites than previously published methods, and it is based on the amplification of a smaller ribosomal DNA fragment. We have tested this protocol in a variety of samples from different geographic regions and various ages of preservation to ascertain the robustness of this protocol over a wide geographic window and on DNA templates of poor quality. This procedure is as efficient as previous ones in discriminating An. arabiensis from the two taxa in An gambiae s.s. However, it performs better than others on poor quality templates such as the ones from museum collections, and poorly stored field collected material. However, it must be noted that it does not allow the simultaneous discrimination of all the species in the An. gambiae complex.

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