Jannotti-Passos Liana K, Magalhães Kelly Grace, Carvalho Omar S, Vidigal Teofânia H D A
Centro de Pesquisas René Rachou-Fiocruz, Av. Augusto de Lima 1715, 30190-002 Belo Horizonte, MG, Brazil.
J Parasitol. 2006 Apr;92(2):401-3. doi: 10.1645/GE-593R1.1.
A simple and single-step technique based on multiplex PCR (multiplex polymerase chain reaction) has been developed for simultaneous identification of Brazilian Biomphalaria species, the intermediate hosts of Schistosoma mansoni, and their diagnosis of infection by the trematode. We used species-specific primers directed both to the internal transcribed spacer 2 (ITS2) of ribosomal DNA from 3 of the S. mansoni host species and to the mitochondrial DNA (mtDNA) from the trematode. Those primers were used simultaneously in a single multiplex-PCR reaction, and template DNA was obtained from S. mansoni-infected and noninfected snails. The results were visualized in silver stained polyacrylamide gels, revealing the presence of specific bands. The methodology has shown to be efficient, fast, and reproducible for Biomphalaria species identification and diagnosis of snails infected by S. mansoni during prepatent periods.
一种基于多重PCR（多重聚合酶链反应）的简单单步技术已被开发出来，用于同时鉴定曼氏血吸虫的中间宿主——巴西双脐螺物种，并诊断其是否感染该吸虫。我们使用了物种特异性引物，这些引物既针对3种曼氏血吸虫宿主物种核糖体DNA的内转录间隔区2（ITS2），也针对该吸虫的线粒体DNA（mtDNA）。这些引物在单个多重PCR反应中同时使用，模板DNA取自感染和未感染曼氏血吸虫的蜗牛。结果在银染聚丙烯酰胺凝胶中可视化，显示出特定条带的存在。该方法已被证明在潜伏期对双脐螺物种鉴定和诊断感染曼氏血吸虫的蜗牛方面是高效、快速且可重复的。
