Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University, Rangsit Campus, 99 Moo 18, Pathumthani 12121, Thailand.
Acta Trop. 2010 May;114(2):76-80. doi: 10.1016/j.actatropica.2010.01.007. Epub 2010 Jan 21.
The liver fluke Opisthorchis viverrini is the agent of human opisthorchiasis in Thailand with a high prevalence observed in the rural population of north and northeastern regions of the country. A focus of research has therefore been the development of diagnostic tools to indicate infection by this parasite. In the present study, a 28 kDa glutathione S-transferase of O. viverrini (OV28GST), which is found in the excretion/secretion product of the parasite, was evaluated for its application in diagnosis of human opisthorchiasis. Bacterially expressed and functionally active rOV28GST was used in immunoblots and indirect ELISA to detect anti-OV28GST antibody in sera of infected individuals. Crude whole worm extract, sera of uninfected individuals and a rabbit anti-rOV28GST antiserum were used as controls in the assays while positivity for parasite DNA by PCR and egg count in faeces were used as primary indicators of infection. The results showed weak or absent reactivity of the infected sera to immunoblotted rOV28GST and no significant difference in absorbance values when compared to uninfected sera in ELISA. In addition, a glutathione capture ELISA which was performed to test for circulating OV28GST in human and hamster sera showed negative results. In conclusion, OV28GST is not applicable as a diagnostic tool in established infections due to low specific antibody titre and abundance as circulating antigen.
肝片形吸虫(Opisthorchis viverrini)是泰国人类华支睾吸虫病的病原体,在该国北部和东北部农村地区的人群中,这种寄生虫的感染率很高。因此,研究的重点是开发诊断工具来指示这种寄生虫的感染。在本研究中,评估了肝片形吸虫的一种 28 kDa 谷胱甘肽 S-转移酶(OV28GST),该酶存在于寄生虫的排泄/分泌产物中,用于诊断人类华支睾吸虫病。在免疫印迹和间接 ELISA 中,使用细菌表达和功能活性的 rOV28GST 来检测感染个体血清中的抗 OV28GST 抗体。在这些检测中,使用粗全虫提取物、未感染个体的血清和兔抗 rOV28GST 抗血清作为对照,而寄生虫 DNA 的 PCR 阳性和粪便中的卵计数则作为感染的主要指标。结果显示,感染血清对免疫印迹 rOV28GST 的反应较弱或不存在,并且在 ELISA 中与未感染血清相比,吸光度值没有显著差异。此外,进行了谷胱甘肽捕获 ELISA 以测试人血清和仓鼠血清中的循环 OV28GST,但结果为阴性。总之,由于特异性抗体滴度低和作为循环抗原的丰度低,OV28GST 不适用于已建立的感染的诊断工具。