Department of Parasitology and Mycology, School of Medicine, Alborz University of Medical Sciences, P.O. Box: 3149779453, Karaj, Iran.
Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Sci Rep. 2024 Feb 16;14(1):3865. doi: 10.1038/s41598-024-54602-y.
Fascioliasis is a zoonotic parasitic infection caused by Fasciola species in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validating methods for the diagnosis of fascioliasis in humans. Serological techniques are convenient assays that significantly improves the diagnosis of Fasciola infection. However, a more sensitive method is required. The aim of this study was to compare the Real-Time PCR technique with the indirect-ELISA for the detection of Fasciola hepatica in human. Using a panel of sera from patients infected with Fasciola hepatica (n = 51), other parasitic infections (n = 7), and uninfected controls (n = 12), we optimized an ELISA which employs an excretory-secretory antigens from F. hepatica for the detection of human fascioliasis. After DNA extraction from the samples, molecular analysis was done using Real-Time PCR technique based on the Fasciola ribosomal ITS1 sequence. Of 70 patient serum samples, 44 (62.86%) samples were identified as positive F. hepatica infection using ELISA and Real-Time PCR assays. There was no cross-reaction with other parasitic diseases such as toxoplasmosis, leishmaniasis, taeniasis, hydatidosis, trichinosis, toxocariasis, and strongyloidiasis. The significant difference between the agreement and similarity of the results of patients with indirect ELISA and Real-Time PCR was 94.4% and 99.2%, respectively (Cohen's kappa ≥ 0.7; P = 0.02). Based on the Kappa agreement findings, the significant agreement between the results of ELISA and Real-Time PCR indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in humans.
片形吸虫病是一种由人类和动物的片形科物种引起的人畜共患寄生虫感染。尽管在疫苗接种和新的治疗药物方面取得了重大进展，但人们很少关注验证人类片形吸虫病诊断方法。血清学技术是一种方便的检测方法，可显著提高对 Fasciola 感染的诊断。然而，需要一种更敏感的方法。本研究旨在比较实时 PCR 技术与间接 ELISA 检测人肝片吸虫的方法。使用一组来自感染肝片吸虫的患者（n=51）、其他寄生虫感染（n=7）和未感染对照（n=12）的血清，我们优化了一种 ELISA，该 ELISA 采用肝片吸虫的排泄分泌抗原检测人类片形吸虫病。从样本中提取 DNA 后，使用基于 Fasciola 核糖体 ITS1 序列的实时 PCR 技术进行分子分析。在 70 份患者血清样本中，44 份（62.86%）样本通过 ELISA 和实时 PCR 检测被鉴定为肝片吸虫感染阳性。与其他寄生虫病（如弓形虫病、利什曼病、带绦虫病、包虫病、旋毛虫病、弓蛔虫病和类圆线虫病）无交叉反应。间接 ELISA 和实时 PCR 检测结果的一致性和相似性的显著差异分别为 94.4%和 99.2%（Cohen's kappa≥0.7；P=0.02）。根据 Kappa 一致性发现，ELISA 和实时 PCR 检测结果之间的显著一致性表明这些检测方法在诊断人类肝片吸虫病中的准确性和可靠性。
