Laboratory of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, Japan.
Acta Trop. 2012 Feb;121(2):93-8. doi: 10.1016/j.actatropica.2011.10.004. Epub 2011 Oct 12.
Genotyping of sand fly species circulating in Peru was established on the basis of PCR-restriction fragment length polymorphisms (RFLPs) of the 18S ribosomal RNA (rRNA) gene. The sequences of 18S rRNA gene fragments from 12 Lutzomyia and 1 Warileya species were determined and their RFLP-patterns were analyzed. Consequently, RFLP analysis with the restriction enzyme AfaI and then HapII or KpnI, followed by XspI successfully differentiated them. Intraspecific genetic diversity affecting RFLP-patterns was not detected in the specimens collected from 24 areas of 8 departments. The genotyping was applied to the surveillance of sand flies collected from Andean areas where leishmaniasis is endemic, and its usability was verified. The present method promises to be a powerful tool for the classification and surveillance of sand flies circulating in Peru.
基于聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析 18S 核糖体 RNA（rRNA）基因，对秘鲁地区的按蚊种类进行基因分型。测定了 12 种卢氏疟蚊和 1 种威拉雷亚蚊的 18S rRNA 基因片段序列，并分析了它们的 RFLP 图谱。结果表明，用 AfaI 内切酶进行 RFLP 分析，然后用 HapII 或 KpnI 以及 XspI 内切酶进行后续处理，可成功地对其进行区分。在从 8 个省的 24 个地区采集的样本中未发现影响 RFLP 图谱的种内遗传多样性。该基因分型方法已应用于对利什曼病流行的安第斯地区采集的按蚊进行监测，并验证了其可用性。本方法有望成为一种强大的工具，用于对秘鲁地区流行的按蚊进行分类和监测。
