Fong Mun Yik, Lau Yee Ling, Zulqarnain Mohamed
Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia.
Biotechnol Lett. 2008 Apr;30(4):611-8. doi: 10.1007/s10529-007-9609-x. Epub 2007 Nov 28.
The surface antigen 2 (SAG2) gene of the protozoan parasite, Toxoplasma gondii, was cloned and extracellularly expressed in the yeast Pichia pastoris. The effectiveness of the secreted recombinant SAG2 (rSAG2-S) as a serodiagnosis reagent was assessed by western blots and ELISA. In the western blot assay, rSAG2-S reacted with all Toxoplasma-antibody positive human serum samples but not with Toxoplasma-negative samples. In the ELISA, rSAG2-S yielded sensitivity rates ranging from 80% (IgG negative, IgM positive) to 100% (IgG positive, IgM negative). In vivo experiments showed that serum from mice immunized with rSAG2-S reacted specifically with the native SAG2 of T. gondii. These mice were protected when challenged with live cells of T. gondii.
对原生动物寄生虫刚地弓形虫的表面抗原2(SAG2)基因进行了克隆,并在酵母毕赤酵母中进行细胞外表达。通过蛋白质印迹法和酶联免疫吸附测定(ELISA)评估分泌型重组SAG2(rSAG2-S)作为血清学诊断试剂的有效性。在蛋白质印迹分析中,rSAG2-S与所有弓形虫抗体阳性的人血清样本发生反应,但不与弓形虫阴性样本反应。在ELISA中,rSAG2-S的灵敏度率范围为80%(IgG阴性,IgM阳性)至100%(IgG阳性,IgM阴性)。体内实验表明,用rSAG2-S免疫的小鼠血清与刚地弓形虫的天然SAG2发生特异性反应。当用刚地弓形虫活细胞攻击这些小鼠时,它们受到了保护。