Laboratory of Veterinary Parasitology, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka 020-8550, Japan.
Parasitol Res. 2010 Feb;106(3):757-61. doi: 10.1007/s00436-010-1724-2. Epub 2010 Jan 28.
Molecular characterization is important for discriminating Fasciola specimens having the deoxyribonucleic acid (DNA) sequences of Fasciola hepatica, Fasciola gigantica, and both Fasciola species, since three Fasciola forms coexist in Asian countries. We have developed a restriction fragment length polymorphism of amplified DNA (PCR-RFLP) of the nuclear ribosomal internal transcribed spacer 1 (ITS1) region in Fasciola species. The band patterns of the fragments digested with a restriction enzyme, Rsa I, were accurately distinguished among the three forms of Fasciola. Amplicons with the sequences of F. hepatica and F. gigantica were divided into fragments of about 360, 100, and 60 bp, and 360, 170, and 60 bp, respectively, and amplicons with the sequences of both Fasciola species yielded fragments of 360, 170, 100, and 60 bp. The results of PCR-RFLP completely coincided with those of sequence analysis, and thus PCR-RFLP is a useful technique for determining the ITS1 type in Fasciola species.
分子特征对于区分具有肝片形吸虫、巨片形吸虫和两种片形吸虫 DNA 序列的片形吸虫标本很重要，因为亚洲国家同时存在三种片形吸虫。我们已经开发了一种针对核核糖体内部转录间隔区 1（ITS1）区域的扩增 DNA（PCR-RFLP）的限制片段长度多态性的方法，用于区分片形吸虫物种。用限制酶 Rsa I 消化片段的带型可以准确地区分三种形式的片形吸虫。来自肝片形吸虫和巨片形吸虫的序列的扩增子被分为约 360、100 和 60 bp 的片段，和 360、170 和 60 bp 的片段，而来自两种片形吸虫物种的序列的扩增子产生 360、170、100 和 60 bp 的片段。PCR-RFLP 的结果与序列分析完全一致，因此 PCR-RFLP 是一种用于确定 ITS1 型片形吸虫的有用技术。
