O'Neill S M, Parkinson M, Dowd A J, Strauss W, Angles R, Dalton J P
School of Biological Sciences, Dublin City University, Ireland.
Am J Trop Med Hyg. 1999 May;60(5):749-51. doi: 10.4269/ajtmh.1999.60.749.
Our laboratory recently developed a diagnostic test (ELISA) for human fascioliasis based on the detection of serum IgG4 antibodies reactive with Fasciola hepatica cathepsin L1 (CL1). In the present study, we have used recombinant CL1, generated by functional expression of the cDNA in Saccharomyces cerevisiae, in this immunodiagnostic test and compared its performance with native CL1. Sera obtained from 64 individuals living in Cutusuma village in the northern Altiplano of Bolivia, a region with a high prevalence of human fascioliasis, were analyzed by the IgG4-ELISA. A highly statistically significant correlation (r2 = 0.751, P < 0.001) was demonstrated between the absorbances obtained using the recombinant and native proteins. These assays showed that 38 (59%) of the individuals tested were seropositive for fascioliasis, whereas only 26 of them were coprologically positive for F. hepatica eggs. All seronegative patients were also coprologically negative. Serum from individuals infected with schistosomiasis mansoni, cysticercosis, hydatidosis, and Chagas disease did not contain antibodies reactive with the recombinant or native CL1. Therefore, recombinant CL1 shows excellent potential for the development of the first standardized assay for the sensitive and specific diagnosis of human fascioliasis. Finally, our data supports earlier reports on the high prevalence of human fascioliasis in the Bolivian Altiplano, which collectively suggest that the disease has been endemic there for more than a decade.
我们实验室最近开发了一种用于人类肝片吸虫病的诊断测试（酶联免疫吸附测定，ELISA），该测试基于检测与肝片吸虫组织蛋白酶L1（CL1）反应的血清IgG4抗体。在本研究中，我们将通过在酿酒酵母中功能性表达cDNA产生的重组CL1用于这种免疫诊断测试，并将其性能与天然CL1进行比较。通过IgG4-ELISA分析了从玻利维亚高原北部库图苏马村的64个人获得的血清，该地区人类肝片吸虫病患病率很高。使用重组蛋白和天然蛋白获得的吸光度之间显示出高度统计学显著相关性（r2 = 0.751，P < 0.001）。这些检测表明，38名（59%）受测个体肝片吸虫病血清学呈阳性，而其中只有26人粪便中肝片吸虫卵检测呈阳性。所有血清学阴性的患者粪便检测也为阴性。感染曼氏血吸虫病、囊尾蚴病、包虫病和恰加斯病的个体血清中不含有与重组或天然CL1反应的抗体。因此，重组CL1在开发首个用于人类肝片吸虫病敏感和特异性诊断的标准化检测方法方面显示出极好的潜力。最后，我们的数据支持了关于玻利维亚高原人类肝片吸虫病高患病率的早期报告，这些报告共同表明该疾病在那里已经流行了十多年。
