Matson Ryan, Rios Carlos Tong, Chavez Cesar Banda, Gilman Robert H, Florin David, Sifuentes Victor Lopez, Greffa Roldan Cardenas, Yori Pablo Peñataro, Fernandez Roberto, Portocarrero Daniel Velasquez, Vinetz Joseph M, Kosek Margaret
University of California San Diego, La Jolla, California 92093, USA.
Am J Trop Med Hyg. 2008 Mar;78(3):492-8.
We evaluated a PCR-RFLP of the ribosomal internal transcribed spacer 2 region (ITS2) to distinguish species of Anopheles commonly reported in the Amazon and validated this method using reared F1 offspring. The following species of Anopheles were used for molecular analysis: An. (Nys.) benarrochi, An. (Nys.) darlingi, An. (Nys.) nuneztovari, An. (Nys.) konderi, An. (Nys.) rangeli, and An. (Nys.) triannulatus sensu lato (s.l.). In addition, three species of the subgenus Anopheles, An. (Ano.) forattini, An. (Ano.) mattogrossensis, and An. (Ano.) peryassui were included for testing. Each of the nine species tested yielded diagnostic banding patterns. The PCR-RFLP method was successful in identifying all life stages including exuviae with small fractions of the sample. The assay is rapid and can be applied as an unbiased confirmatory method for identification of morphologic variants, disputed samples, imperfectly preserved specimens, and life stages from which taxonomic keys do not allow for definitive species determination.
我们评估了核糖体内部转录间隔区2（ITS2）区域的聚合酶链反应-限制性片段长度多态性（PCR-RFLP），以区分亚马逊地区常见的按蚊种类，并使用饲养的F1后代验证了该方法。以下按蚊种类用于分子分析：贝氏按蚊（An. (Nys.) benarrochi）、达氏按蚊（An. (Nys.) darlingi）、努氏按蚊（An. (Nys.) nuneztovari）、康德按蚊（An. (Nys.) konderi）、兰氏按蚊（An. (Nys.) rangeli）和广义三带按蚊（An. (Nys.) triannulatus sensu lato (s.l.)）。此外，还纳入了按蚊亚属的三种按蚊进行测试，即福氏按蚊（An. (Ano.) forattini）、马托格罗索按蚊（An. (Ano.) mattogrossensis）和佩氏按蚊（An. (Ano.) peryassui）。所测试的九种按蚊均产生了诊断性条带模式。PCR-RFLP方法成功地鉴定了所有生命阶段，包括仅需少量样本的蜕皮。该检测方法快速，可作为一种无偏倚的确认方法，用于鉴定形态变异体、有争议的样本、保存不完美的标本以及分类检索表无法确定其确切种类的生命阶段。
