Molecular Parasitology Unit, Laboratory of Parasitic Diseases, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0425, USA.
Vet Parasitol. 2010 Apr 19;169(1-2):37-44. doi: 10.1016/j.vetpar.2009.12.020. Epub 2009 Dec 22.
Sarcocystis neurona is an apicomplexan parasite identified as a cause of fatal neurological disease in the threatened southern sea otter (Enhydra lutris nereis). In an effort to characterize virulent S. neurona strains circulating in the marine ecosystem, this study developed a range of markers relevant for molecular genotyping. Highly conserved sequences within the 18S ribosomal gene array, the plastid-encoded RNA polymerase (RPOb) and the cytochrome c oxidase subunit 1 mitochondrial gene (CO1) were assessed for their ability to distinguish isolates at the genus and species level. For within-species comparisons, five surface antigens (SnSAG1-SnSAG5) and one high resolution microsatellite marker (Sn9) were developed as genotyping markers to evaluate intra-strain diversity. Molecular analysis at multiple loci revealed insufficient genetic diversity to distinguish terrestrial isolates from strains infecting marine mammals. Furthermore, SnSAG specific primers applied against DNA from the closely related species, Sarcocystis falcatula, lead to the discovery of highly similar orthologs to SnSAG2, 3, and 4, calling into question the specificity of diagnostic tests based on these antigens. The results of this study suggest a population genetic structure for S. neurona similar to that reported for the related parasite, Toxoplasma gondii, dominated by a limited number of successful genotypes.
神经内罗巴原虫(Sarcocystis neurona)是一种顶复门寄生虫,被认为是濒危的南方海獭(Enhydra lutris nereis)致命神经疾病的病因。为了描述在海洋生态系统中循环的毒力 S. neurona 菌株,本研究开发了一系列与分子基因分型相关的标记物。评估了 18S 核糖体基因阵列、质体编码的 RNA 聚合酶(RPOb)和细胞色素 c 氧化酶亚基 1 线粒体基因(CO1)内的高度保守序列,以确定其在属和种水平上区分分离株的能力。对于种内比较,开发了五个表面抗原(SnSAG1-SnSAG5)和一个高分辨率微卫星标记(Sn9)作为基因分型标记,以评估菌株内的多样性。多个位点的分子分析表明,遗传多样性不足以区分来自陆地的分离株和感染海洋哺乳动物的菌株。此外,针对与 S. neurona 密切相关的物种 Sarcocystis falcatula 的 DNA 应用 SnSAG 特异性引物,导致发现与 SnSAG2、3 和 4 高度相似的直系同源物,这使得基于这些抗原的诊断测试的特异性受到质疑。本研究的结果表明,S. neurona 的群体遗传结构与相关寄生虫弓形虫(Toxoplasma gondii)相似,主要由少数成功的基因型主导。