Department of Parasitology, West China School of Preclinical and Forensic Medicine, Sichuan University, No. 17 Section 3, Renmin South Road, Chengdu 610041, Sichuan, China.
Vet Parasitol. 2012 Feb 10;183(3-4):353-5. doi: 10.1016/j.vetpar.2011.07.044. Epub 2011 Aug 4.
This work describes a simple method to yield large amounts of the isolate MHOM/CN/90/SC10H2 amastigotes-like forms in axenic cultures using promastigotes as the starting population. The isolate MHOM/CN/90/SC10H2, used in this study, belongs to an undescribed species of Leishmania endemic to hill foci in China. The method describes induced extracellular amastigote transformation of this isolate. The rounded parasite obtained in axenic culture was morphologically similar, even at the ultrastructural level, to intracellular amastigotes. Moreover, the axenic amastigotes remained viable as verified by the stage-specific genes (gp46 and p4 genes) with RT-PCR. A 70-80 kDa protein was recognized by polyclonal antibody HRP-IgG only in axenic-derived amastigotes and not in promastigotes.
本工作描述了一种简单的方法，可使用无鞭毛体作为起始群体，在无共生菌培养物中大量产生 MHOM/CN/90/SC10H2 分离株的前鞭毛体样形式。本研究中使用的 MHOM/CN/90/SC10H2 分离株属于中国丘陵热点地区特有的未描述的利什曼原虫种。该方法描述了该分离株的诱导细胞外无鞭毛体转化。在无共生菌培养物中获得的圆形寄生虫在形态上甚至在超微结构水平上都与细胞内无鞭毛体相似。此外，通过 RT-PCR 对阶段特异性基因（gp46 和 p4 基因）的验证，证明无共生菌来源的无鞭毛体仍然具有活力。多克隆抗体 HRP-IgG 仅在无共生菌衍生的无鞭毛体中识别 70-80 kDa 蛋白，而不在前鞭毛体中识别。
