Mathis A, Akerstedt J, Tharaldsen J, Odegaard O, Deplazes P
Institute of Parasitology, CH-8057 Zürich, Switzerland.
Parasitol Res. 1996;82(8):727-30. doi: 10.1007/s004360050192.
Encephalitozoon cuniculi has a wide host range among mammals, but whether it represents a homogeneous species is a subject of controversy. We have isolated, cultivated (in human MRC-5 cells) and, for the first time, characterized by immunological and molecular biological methods four isolates of E. cuniculi from Norwegian blue foxes with a history of encephalitozoonosis. The isolates were compared with nine isolates from domestic rabbits from Switzerland. Two E. cuniculi subtypes were identified according to their host species. A 5'-GTTT-3' tetranucleotide repeat was present twice in the rDNA intergenic spacer in all isolates from foxes as opposed to three times in all isolates from rabbits. Furthermore, random amplified polymorphic DNA analysis showed one polymorphic band among the subtypes, and Western-blot analysis using serum from an infected fox discriminated between the two subtypes on the basis of their banding patterns in the ranges of 31-33 and 38-40 kDa. The 5'-GTTT-3' tetranucleotide repeat is a valuable genetic marker for these two subtypes of E. cuniculi and will be of use in continued studies on the molecular epidemiology of this parasite.
兔脑炎微孢子虫在哺乳动物中有广泛的宿主范围，但它是否代表一个同质物种是一个有争议的问题。我们从患有脑炎微孢子虫病的挪威蓝狐中分离、培养（在人MRC-5细胞中）并首次通过免疫和分子生物学方法鉴定了四株兔脑炎微孢子虫。将这些分离株与来自瑞士家兔的九株分离株进行了比较。根据宿主物种鉴定出两种兔脑炎微孢子虫亚型。在所有来自狐狸的分离株中，rDNA基因间隔区存在两次5'-GTTT-3'四核苷酸重复，而在所有来自兔子的分离株中存在三次。此外，随机扩增多态性DNA分析显示各亚型之间有一条多态带，使用感染狐狸的血清进行的蛋白质免疫印迹分析根据其在31-33 kDa和38-40 kDa范围内的条带模式区分了这两种亚型。5'-GTTT-3'四核苷酸重复是这两种兔脑炎微孢子虫亚型的一个有价值的遗传标记，将用于对该寄生虫分子流行病学的后续研究。
