Schmatz D M, Crane M S, Murray P K
J Protozool. 1986 Feb;33(1):109-14. doi: 10.1111/j.1550-7408.1986.tb05568.x.
An assay has been developed using parasite-specific incorporation of 3H-uracil to assess the intracellular growth of Eimeria tenella in vitro. As shown by both scintillation counts and autoradiography, 3H-uracil was incorporated specifically into intracellular parasites from the onset of infection and continued throughout development of the first generation schizonts. Mature schizonts and first generation merozoites did not continue to incorporate additional 3H-uracil, indicating that RNA synthesis had halted in these stages. Based on these findings, a semi-automated microscale uracil incorporation assay was developed to determine parasite viability. This method should be useful for biochemical studies with intracellular parasites and for screening compounds for anticoccidial activity. The ease, rapidity, and quantitative nature of this assay contrasts favorably with standard morphometric approaches of determining parasite development. In addition, parallel studies using host cell incorporation of 3H-uridine have been introduced as a method of determining whether antiparasitic activity is direct or indirect in relation to effects on the host cell.
已开发出一种利用寄生虫特异性掺入3H-尿嘧啶来评估柔嫩艾美耳球虫体外细胞内生长的检测方法。如闪烁计数和放射自显影所示，从感染开始，3H-尿嘧啶就特异性地掺入细胞内寄生虫中，并在第一代裂殖体的整个发育过程中持续存在。成熟裂殖体和第一代裂殖子不再继续掺入额外的3H-尿嘧啶，这表明在这些阶段RNA合成已经停止。基于这些发现，开发了一种半自动微量尿嘧啶掺入检测方法来确定寄生虫的活力。该方法对于细胞内寄生虫的生化研究以及筛选抗球虫活性化合物应该是有用的。该检测方法的简便、快速和定量性质与确定寄生虫发育的标准形态计量学方法相比具有优势。此外，已引入使用宿主细胞掺入3H-尿苷的平行研究，作为确定抗寄生虫活性相对于对宿主细胞的影响是直接还是间接的一种方法。
