Ramachandran S, Gam A A, Neva F A
Clinical Parasitology Unit, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Am J Trop Med Hyg. 1997 Jan;56(1):61-5. doi: 10.4269/ajtmh.1997.56.61.
The relationships between the parasitic nematodes of medical importance belonging to the genus Strongyloides was studied using a polymerase chain reaction (PCR)-linked restriction fragment length polymorphism approach. We used several human and dog isolates of S. stercoralis, a monkey isolate of S. fulleborni, and S. ratti, a rodent parasite. The molecular analysis was based on amplification of the internal transcribed spacer and the 5' portion of the 23S-like rRNA gene followed by restriction enzyme digests. The length of the PCR product was specific to each species and varied around 1.5 kilobase pairs. Using nine restriction enzymes, we were able to analyze both interspecific and intraspecific variations. With four restriction enzymes (Taq I, Dde I, Dra I, and Mwo I), human isolates of S. stercoralis from different parts of the world showed identical patterns and could be differentiated from the dog isolate of S. stercoralis. Interspecific differences were readily observed with these and other enzymes. In addition to providing new information on the genomic characteristics of Strongyloides parasites, the results suggest that this technique could be useful for diagnostic and epidemiologic investigations.
利用聚合酶链反应(PCR)关联的限制性片段长度多态性方法,对粪类圆线虫属具有医学重要性的寄生线虫之间的关系进行了研究。我们使用了几种人源和犬源的粪类圆线虫分离株、一种来自猴子的富氏类圆线虫分离株以及一种啮齿动物寄生虫——鼠类圆线虫。分子分析基于对内部转录间隔区和23S样rRNA基因5'部分的扩增,随后进行限制性酶切。PCR产物的长度对每个物种而言是特定的,约为1.5千碱基对。使用九种限制性酶,我们能够分析种间和种内变异。使用四种限制性酶(Taq I、Dde I、Dra I和Mwo I)时,来自世界不同地区的人源粪类圆线虫分离株呈现出相同的模式,并且能够与犬源粪类圆线虫分离株区分开来。利用这些酶和其他酶很容易观察到种间差异。这些结果除了提供关于类圆线虫寄生虫基因组特征的新信息外,还表明该技术可用于诊断和流行病学调查。