de Leeuw W A, Cornelissen J B
Central Veterinary Institute, Department of Immunology, Lelystad, Netherlands.
Vet Parasitol. 1993 Sep;49(2-4):229-41. doi: 10.1016/0304-4017(93)90122-4.
Three enzyme-linked immunosorbent assays (ELISA) that detect antibodies against Dictyocaulus viviparus in cattle were compared for sensitivity, specificity and seroconversion after primary infection. These assays were (i) an indirect ELISA with crude somatic antigens from adult D. viviparus (ca-ELISA), (ii) an indirect ELISA with purified antigens (sa-ELISA) isolated from somatic antigens of adult D. viviparus and (iii) a competition ELISA with antigen purified with anion chromatography in combination with monoclonal antibodies against D. viviparus. Sera from helminth-naïve calves and sera from calves monospecifically infected with Ostertagia ostertagi, Cooperia oncophora, Nematodirus helvetianus, Ascaris suum or Fasciola hepatica were used to determine the specificity of the assays. Sera from calves and milk cows experimentally or naturally infected with D. viviparus, and from vaccinated calves, were used to test the sensitivity of the assays and to determine when the ELISAs detected seroconversion. The specificity of the competition and the sa-ELISA was 97%, whereas the specificity of the ca-ELISA was 67%. The sensitivities of the sa-ELISA, the competition ELISA and the ca-ELISA were 97, 73 and 99%, respectively. All three assays detected seroconversion between 4 and 6 weeks after primary infection. None of the assays detected seroconversion in calves receiving lungworm vaccination. We conclude that of these three tests, the sa-ELISA can be used most beneficially to diagnose lungworm disease.
对三种检测牛体内抗胎生网尾线虫抗体的酶联免疫吸附测定(ELISA)方法进行了比较,以评估其在初次感染后的敏感性、特异性和血清转化情况。这三种方法分别为:(i)使用成年胎生网尾线虫粗体抗原的间接ELISA(ca-ELISA);(ii)使用从成年胎生网尾线虫体抗原中分离出的纯化抗原的间接ELISA(sa-ELISA);(iii)将阴离子色谱法纯化的抗原与抗胎生网尾线虫单克隆抗体结合使用的竞争ELISA。使用未感染蠕虫的犊牛血清以及单特异性感染奥斯特他线虫、柯氏尖尾线虫、瑞士细颈线虫、猪蛔虫或肝片吸虫的犊牛血清来确定这些检测方法的特异性。使用经实验或自然感染胎生网尾线虫的犊牛和奶牛血清以及接种疫苗的犊牛血清来测试这些检测方法的敏感性,并确定ELISA检测到血清转化的时间。竞争ELISA和sa-ELISA的特异性为97%,而ca-ELISA的特异性为67%。sa-ELISA、竞争ELISA和ca-ELISA的敏感性分别为97%、73%和99%。所有三种检测方法均在初次感染后4至6周检测到血清转化。在接受肺线虫疫苗接种的犊牛中,没有一种检测方法检测到血清转化。我们得出结论,在这三种检测方法中,sa-ELISA最有利于诊断肺线虫病。