Department of Microbiology, University of Tennessee, Knoxville, TN, USA.
Agricultural Research Service, Animal Parasitic Diseases Laboratory, United States Department of Agriculture (USDA), Beltsville Agricultural Research Center, Beltsville, MD, USA.
Methods Mol Biol. 2020;2071:49-80. doi: 10.1007/978-1-4939-9857-9_3.
Toxoplasma gondii is a protozoan parasite that infects mammals and birds. Molecular epidemiology and population genetic studies have revealed widespread and distinct distribution of different T. gondii genotypes globally. Animals (domestic and wild) are the reservoirs for transmission of this parasite to humans. Recent development in molecular genotyping methods allowed us to identify parasite strains with high resolution and to dissect transmission patterns among different hosts. However, current data in the literature is still limited and fragmented. Here, we summarize a set of protocols that can be used to identify T. gondii infection in clinically normal animals, isolate the parasite by bioassay using animal tissues, extract parasite DNA from tissue samples, and finally identify the parasite by multilocus PCR-RFLP genotyping. We hope these protocols provide essential tools to study genetic diversity, population structure and transmission dynamics of T. gondii. Accumulation of the information will allow us to better understand, control, and prevent T. gondii infection in the future.
刚地弓形虫是一种感染哺乳动物和鸟类的原生动物寄生虫。分子流行病学和种群遗传学研究揭示了全球不同刚地弓形虫基因型的广泛而独特的分布。动物(家养和野生)是该寄生虫向人类传播的储存宿主。分子基因分型方法的最新进展使我们能够以高分辨率识别寄生虫株,并剖析不同宿主之间的传播模式。然而,目前文献中的数据仍然有限且分散。在这里,我们总结了一套方案,可用于鉴定临床正常动物中的刚地弓形虫感染,通过动物组织的生物测定法分离寄生虫,从组织样本中提取寄生虫 DNA,最后通过多位点 PCR-RFLP 基因分型鉴定寄生虫。我们希望这些方案为研究刚地弓形虫的遗传多样性、种群结构和传播动力学提供必要的工具。信息的积累将使我们能够更好地理解、控制和预防未来的刚地弓形虫感染。