Discrimination of all members of the Anopheles punctulatus complex by polymerase chain reaction--restriction fragment length polymorphism analysis.
A method has been developed to identify the members of the Anopheles punctulatus complex using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Members of the An. punctulatus complex are the most important vectors of malaria in the southwest Pacific and consist of 10 cryptic species, An. farauti no. 1-7, An. punctulatus, An. sp. near punctulatus, and An. koliensis. For each species, PCR amplification of the ribosomal DNA internal transcribed spacer produced a 750-basepair product. Digestion with Msp I and electrophoresis on a 3.0% agarose gel results in banding patterns unique to each species. Isolates of the same species from different locations gave an identical pattern. The technique is sensitive enough so that a PCR-RFLP can be generated from as little as a single mosquito leg, allowing the rest of the mosquito to be used for other important epidemiologic analyses such as determining host feeding source, and for parasite detection.
D000818:Animals; D000852:Anopheles / Q000145:classification / Q000235:genetics*; D001315:Australia; D017931:DNA Primers / Q000737:chemistry; D004275:DNA, Ribosomal / Q000032:analysis / Q000737:chemistry; D007303:Insect Vectors / Q000145:classification / Q000235:genetics*; D008542:Melanesia; D010219:Papua New Guinea; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity; D009514:Vanuatu
D017931:DNA Primers; D004275:DNA, Ribosomal