Khodadadi A, Madani R, Hoghooghi Rad N, Atyabi N
Department of Pathobiology, Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Proteomics, Biochemistry and Biotechnology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO) Karaj, Iran.
Arch Razi Inst. 2021 Jan;75(4):419-426. doi: 10.22092/ari.2018.123028.1236. Epub 2021 Jan 1.
Toxoplasmosis is a widespread parasitic disease caused by a protozoan parasite Toxoplasma gondii. Currently, nanotechnology has been used for the diagnosis of many infectious diseases. It could be due to the fact that nanoparticles play an important role in accurate and fast diagnosis. The purpose of this study was to design a Nano-enzyme linked immunosorbent assay (Nano-ELISA) kit using excreted/secreted (E/S) antigens to have higher sensitivity and specificity than those reported for the designed enzyme-linked immunosorbent assay (ELISA) kit for the diagnosis of Toxoplasmosis in mice. Firstly, the serum samples were collected from 15 infected mice with T. gondii and 15 healthy ones. Then, E/S antigens were separated from parasite tachyzoites and used for designing an ELISA kit. In addition, the mice sera were evaluated using the designed ELISA kit. Finally, the serum samples were assessed by Nano-ELISA kits designed with E/S antigen and conjugate of gold nanoparticles. The obtained results of the present study showed that the sensitivity and specificity of the designed ELISA kit were reported as 80% and 86.66%, respectively, that both improved to 93.33% in these sera with the designed Nano-ELISA kit. This finding revealed the significant improvement of sensitivity and specificity using gold nanoparticles in designing the ELISA kit. Furthermore, according to the literature, the use of E/S antigens in designing recognizable ELISA kits has been always highlighted considering the presence of numerous antigens in T. gondii. The results of this study revealed that the use of E/S antigens in the preparation of an ELISA kit was very effective. This is very important, especially in the lower titers of antibody requiring a more accurate diagnosis. On the other hand, the Nano-ELISA method designed with E/S antigens can be more sensitive and specific than ELISA for the diagnosis of Toxoplasmosis and can be the basis for further studies in this regard.
弓形虫病是一种由原生动物寄生虫刚地弓形虫引起的广泛传播的寄生虫病。目前,纳米技术已被用于多种传染病的诊断。这可能是因为纳米颗粒在准确快速诊断中发挥着重要作用。本研究的目的是设计一种使用排泄/分泌(E/S)抗原的纳米酶联免疫吸附测定(Nano-ELISA)试剂盒,使其比已报道的用于诊断小鼠弓形虫病的酶联免疫吸附测定(ELISA)试剂盒具有更高的灵敏度和特异性。首先,从15只感染刚地弓形虫的小鼠和15只健康小鼠中采集血清样本。然后,从寄生虫速殖子中分离出E/S抗原,并用于设计ELISA试剂盒。此外,使用设计好的ELISA试剂盒对小鼠血清进行评估。最后,用含有E/S抗原和金纳米颗粒缀合物的Nano-ELISA试剂盒对血清样本进行评估。本研究获得的结果表明,设计的ELISA试剂盒的灵敏度和特异性分别为80%和86.66%,而使用设计的Nano-ELISA试剂盒时,这些血清中的灵敏度和特异性均提高到了93.33%。这一发现揭示了在设计ELISA试剂盒时使用金纳米颗粒可显著提高灵敏度和特异性。此外,根据文献,考虑到刚地弓形虫中存在大量抗原,在设计可识别的ELISA试剂盒时使用E/S抗原一直受到重视。本研究结果表明,在制备ELISA试剂盒时使用E/S抗原非常有效。这非常重要,尤其是在抗体滴度较低需要更准确诊断的情况下。另一方面,用E/S抗原设计的Nano-ELISA方法在诊断弓形虫病时可能比ELISA更灵敏、更特异,可作为这方面进一步研究的基础。