Yaman Kerem, Akarsu Gülay Aral, Güngör Çiğdem, Ataoğlu Haluk
Ankara Üniversitesi Tıp Fakültesi, Tıbbi Parazitoloji Bilim Dalı, Ankara, Turkey.
Turkiye Parazitol Derg. 2011;35(3):133-6. doi: 10.5152/tpd.2011.33.
Toxoplasma gondii the causative agent of toxoplasmosis, is a worldwide intracellular protozoan parasite that infects all warm blooded animals including humans. It can be devastating to immunocompromised humans and congenital transmission may result in severe clinical spectrum. It causes economic losses due to abortus in animals. Toxoplasmosis diagnosis depends on direct and indirect methods. Besides the Sabin-Feldman test, which is accepted to be the reference test, serologic tests such as ELISA and immunofluorescence antibody tests are means of indirect diagnosis. As detected antibodies in serologic tests are correlated with antigens that cause their synthesis, it is important to know different proteins of different strains. In this study RH, Ankara and TS-4 strains were used and differences between their proteins were examined.
RH and Ankara strains were inoculated into the peritoneal cavity of mice. TS- 4 strain was produced in Vero cell culture. Tachyzoites collected by peritoneal wash were lysed and lyophilised. This was run on SDS-PAGE gel and protein bands were compared with a standard protein ladder after staining with polychromatic silver stain.
It was observed that, while Ankara and RH strains had dense bands between 60-70 kDa and at 15 kDa, the most prominent bands of TS-4 strain were 60 ve 115 kDa bands.
RH and Ankara strains have the same protein bands while TS-4 strain has different and fewer protein bands than the others.
弓形虫是弓形虫病的病原体，是一种全球范围内的细胞内原生动物寄生虫，可感染包括人类在内的所有温血动物。它对免疫功能低下的人类可能是毁灭性的，先天性传播可能导致严重的临床症状。它还会因动物流产而造成经济损失。弓形虫病的诊断依赖于直接和间接方法。除了被公认为参考检测方法的萨宾-费尔德曼试验外，酶联免疫吸附测定（ELISA）和免疫荧光抗体试验等血清学检测是间接诊断手段。由于血清学检测中检测到的抗体与导致其合成的抗原相关，了解不同菌株的不同蛋白质很重要。在本研究中，使用了RH、安卡拉和TS-4菌株，并检测了它们蛋白质之间的差异。
将RH和安卡拉菌株接种到小鼠腹腔中。TS-4菌株在非洲绿猴肾细胞（Vero）培养物中产生。通过腹腔冲洗收集的速殖子被裂解并冻干。将其在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）凝胶上进行电泳，用多色银染染色后，将蛋白条带与标准蛋白梯度进行比较。
观察到，安卡拉和RH菌株在60 - 70 kDa之间和15 kDa处有密集条带，而TS-4菌株最突出的条带是60和115 kDa条带。
RH和安卡拉菌株具有相同的蛋白条带，而TS-4菌株具有与其他菌株不同且更少的蛋白条带。
