Cabán-Hernández Kimberly, Gaudier José F, Ruiz-Jiménez Caleb, Espino Ana M
Laboratory of Immunology and Molecular Parasitology, Department of Microbiology, University of Puerto Rico, School of Medicine, San Juan, Puerto Rico.
J Clin Microbiol. 2014 Mar;52(3):766-72. doi: 10.1128/JCM.02875-13. Epub 2013 Dec 18.
Coprological examination based on egg detection in stool samples is currently used as the gold standard for the diagnosis of human fascioliasis. However, this method is not effective during the acute phase of the disease and has poor sensitivity during the chronic phase. Serodiagnosis has become an excellent alternative to coprological examination in efforts to combat the effects of fascioliasis on human and animal health. Two novel recombinant Fasciola hepatica proteins, i.e., a ferritin (FhFtn-1) and a tegument-associated protein (FhTP16.5), were used as antigens to develop in-house enzyme-linked immunosorbent assay (ELISA) methods. The assays were optimized and validated using 152 serum samples from humans with a known infection status, including healthy subjects, patients with chronic fascioliasis, and patients with other parasitic diseases. The FhFtn-1 ELISA was shown to be 96.6% sensitive and 95.7% specific; the respective parameters for the FhTP16.5 ELISA were 91.4% and 92.4%. The performances of the FhFtn-1 and FhTP16.5 ELISAs were compared with that of an available commercial test (the DRG test) using a subset of serum samples. Our in-house tests were slightly more sensitive than the DRG test in detecting antibodies against F. hepatica, but the differences were not statistically significant. In conclusion, the present study provides evidence for the potential of the FhFtn-1 and FhTP16.5 ELISAs as diagnostic tools for human fascioliasis, as might be implemented in conjunction with standard assays for large-scale screenings in areas where the disease is endemic and for the detection of occasional cases in clinical laboratories.
基于粪便样本中虫卵检测的粪便学检查目前被用作人类肝片吸虫病诊断的金标准。然而，该方法在疾病急性期无效，在慢性期敏感性较差。血清学诊断已成为粪便学检查的一种极佳替代方法，以努力对抗肝片吸虫病对人类和动物健康的影响。两种新型重组肝片吸虫蛋白，即一种铁蛋白（FhFtn-1）和一种皮层相关蛋白（FhTP16.5），被用作抗原，开发了内部酶联免疫吸附测定（ELISA）方法。使用来自已知感染状况的152份人类血清样本（包括健康受试者、慢性肝片吸虫病患者和其他寄生虫病患者）对这些测定进行了优化和验证。FhFtn-1 ELISA的敏感性为96.6%，特异性为95.7%；FhTP16.5 ELISA的相应参数分别为91.4%和92.4%。使用一部分血清样本，将FhFtn-1和FhTP16.5 ELISA的性能与一种现有的商业检测方法（DRG检测）进行了比较。我们的内部检测方法在检测抗肝片吸虫抗体方面比DRG检测略敏感，但差异无统计学意义。总之，本研究为FhFtn-1和FhTP16.5 ELISA作为人类肝片吸虫病诊断工具的潜力提供了证据，可与标准检测方法结合用于疾病流行地区的大规模筛查以及临床实验室中偶发病例的检测。
