Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Exp Parasitol. 2010 Oct;126(2):278-80. doi: 10.1016/j.exppara.2010.05.007. Epub 2010 May 21.
Merozoite invasion of erythrocytes is a crucial step for the asexual cycle of Plasmodium falciparum. Multiple invasion pathways, which involve different ligand-receptor interactions, have been identified in P. falciparum by examining the entry of purified parasite into erythrocytes with different surface receptors, either mutant or under different enzyme treatments. The most critical step for a successful invasion assay is the isolation of erythrocytes infected with viable schizonts. Here, we applied a magnetic column to purify the schizonts for the erythrocyte invasion assay. Comparing to Percoll-sorbitol purification method, this modified approach showed great improvement on reproducibility and reliability of invasion assay, particularly for short-term, culture-adapted parasite isolates. The magnetic purification method is an excellent alternative for parasite isolates that do not tolerate or with unknown sensitivity to Percoll-sorbitol exposure.
裂殖子入侵红细胞是恶性疟原虫无性生殖周期的关键步骤。通过研究纯化的寄生虫与具有不同表面受体的红细胞的结合,包括突变体或不同酶处理的红细胞,已经在恶性疟原虫中鉴定出多种入侵途径,这些途径涉及不同的配体-受体相互作用。成功入侵试验的最关键步骤是分离感染有活裂殖体的红细胞。在这里,我们应用了一个磁柱来纯化裂殖子,以进行红细胞入侵试验。与聚蔗糖-泛影葡胺(Percoll-sorbitol)纯化方法相比,这种改良的方法显著提高了入侵试验的可重复性和可靠性,尤其是对于短期培养适应的寄生虫分离株。对于不能耐受或对聚蔗糖-泛影葡胺暴露的敏感性未知的寄生虫分离株,该磁珠纯化方法是一个很好的替代方法。