Beebe N W, Saul A
Tropical Health Program, Queensland Institute of Medical Research, Brisbane, Australia.
Am J Trop Med Hyg. 1995 Nov;53(5):478-81. doi: 10.4269/ajtmh.1995.53.478.
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.
已开发出一种利用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）来鉴定 punctulatus 按蚊复合体成员的方法。punctulatus 按蚊复合体的成员是西南太平洋地区最重要的疟疾传播媒介，由 10 个隐种组成，即法氏按蚊 1 - 7 号、punctulatus 按蚊、punctulatus 近缘按蚊和 koliensis 按蚊。对于每个物种，核糖体 DNA 内转录间隔区的 PCR 扩增产生一个 750 碱基对的产物。用 Msp I 消化并在 3.0%琼脂糖凝胶上进行电泳，会产生每个物种独特的条带模式。来自不同地点的同一物种分离株呈现相同的模式。该技术灵敏度足够高，以至于仅从一条蚊子腿就可以产生 PCR-RFLP，从而使蚊子的其余部分可用于其他重要的流行病学分析，如确定宿主吸血来源以及进行寄生虫检测。
