使用生物素化抗体的双抗体夹心酶联免疫吸附测定法检测犬细粒棘球绦虫粪抗原

Double-antibody sandwich ELISA using biotinylated antibodies for the detection of Echinococcus granulosus coproantigens in dogs.

作者信息

Benito Aitziber, Carmena David

机构信息

Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy, University of the Basque Country, Vitoria, Spain.

出版信息

Acta Trop. 2005 Jul;95(1):9-15. doi: 10.1016/j.actatropica.2005.03.005.

DOI:10.1016/j.actatropica.2005.03.005

PMID:15863093

Abstract

Here we present the diagnostic evaluation of an improved double-antibody sandwich ELISA for detecting Echinococcus granulosus antigens in dog faecal samples (coproantigens). A purified rabbit IgG fraction against protoscolex excretory-secretory products was used as primary antibody, and the same fraction conjugated with biotin as secondary antibody. In order to validate the sandwich ELISA, intra- and inter-assay precision, linearity, and recovery percentages were calculated. The diagnostic evaluation of the method was carried out by investigating faecal samples from 37 dogs naturally infected with E. granulosus, 15 Echinococcus-free dogs infected with Taenia spp., 82 dogs with non-taeniid helminths and 66 dogs free of helminth infections. An overall sensitivity of 78.4% and specificity of 93.3% were determined. Positive and negative predictive values were 72 and 95%, respectively, and the diagnostic efficiency was 90.5%. In addition, the sandwich ELISA detection limit was estimated in 5.12 ng ml(-1). These results are highly satisfactory, allowing the use of this methodology in surveillance and control programs for intestinal echinococcosis in dogs.

摘要

在此，我们展示了一种改良的双抗体夹心酶联免疫吸附测定法（ELISA）用于检测犬粪便样本（粪抗原）中细粒棘球绦虫抗原的诊断评估。一种针对原头节排泄-分泌产物的纯化兔IgG组分用作一抗，与生物素偶联的相同组分用作二抗。为了验证夹心ELISA，计算了批内和批间精密度、线性以及回收率。通过调查37只自然感染细粒棘球绦虫的犬、15只感染带绦虫属但无棘球绦虫的犬、82只感染非带绦虫类蠕虫的犬以及66只无蠕虫感染的犬的粪便样本，对该方法进行了诊断评估。确定总体敏感性为78.4%，特异性为93.3%。阳性和阴性预测值分别为72%和95%，诊断效率为90.5%。此外，夹心ELISA检测限估计为5.12 ng ml⁻¹。这些结果非常令人满意，使得该方法可用于犬肠道棘球蚴病的监测和控制项目。