Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo SP 05508-000, Brazil.
Vet Parasitol. 2011 Mar 10;176(2-3):275-80. doi: 10.1016/j.vetpar.2010.10.054. Epub 2010 Nov 4.
Coccidiosis are the major parasitic diseases in poultry and other domestic animals including the domestic rabbit (Oryctolagus cuniculus). Eleven distinct Eimeria species have been identified in this host, but no PCR-based method has been developed so far for unequivocal species differentiation. In this work, we describe the development of molecular diagnostic assays that allow for the detection and discrimination of the 11 Eimeria species that infect rabbits. We determined the nucleotide sequences of the ITS1 ribosomal DNAs and designed species-specific primers for each species. We performed specificity tests of the assays using heterologous sets of primers and DNA samples, and no cross-specific bands were observed. We obtained a detection limit varying from 500fg to 1pg, which corresponds approximately to 0.8-1.7 sporulated oocysts, respectively. The test reported here showed good reproducibility and presented a consistent sensitivity with three different brands of amplification enzymes. These novel diagnostic assays will permit population surveys to be performed with high sensitivity and specificity, thus contributing to a better understanding of the epidemiology of this important group of coccidian parasites.
球虫病是家禽和其他家畜（包括家兔（Oryctolagus cuniculus））的主要寄生虫病。在这种宿主中已鉴定出 11 种不同的艾美耳球虫，但迄今为止尚未开发出基于 PCR 的明确物种鉴别方法。在这项工作中，我们描述了分子诊断检测方法的开发，该方法允许检测和区分感染兔子的 11 种艾美耳球虫。我们确定了 ITS1 核糖体 DNA 的核苷酸序列，并为每种物种设计了特异性引物。我们使用异源引物和 DNA 样品进行了检测方法的特异性测试，没有观察到交叉特异性条带。我们获得了从 500fg 到 1pg 的检测限，分别对应于大约 0.8-1.7 个孢子化卵囊。本报告中的检测具有良好的重现性，并与三种不同品牌的扩增酶表现出一致的灵敏度。这些新的诊断检测方法将以高灵敏度和特异性进行种群调查，从而有助于更好地了解这组重要的球虫寄生虫的流行病学。
