Cornelissen J B, Borgsteede F H, van Milligen F J
Department of Immunology, DLO-Institute for Animal Science and Health, Lelystad, Netherlands.
Vet Parasitol. 1997 Jun;70(1-3):153-64. doi: 10.1016/s0304-4017(96)01141-7.
An enzyme-linked immunosorbent assay (ELISA) that detects antibodies against Dictyocaulus viviparus in experimentally and naturally infected cattle was evaluated for its sensitivity, specificity, the moment of seroconversion and persistence of the anti-D. viviparus response and precision. The first three parameters were compared with those of an indirect haemagglutination assay (IHA). Specificity and sensitivity of both assays were assessed in sera collected from calves experimentally infected with pure isolates of D. viviparus, Ostertagia ostertagi, Cooperia oncophora, Nematodirus helvetianus, Ascaris suum or Fasciola hepatica, and from parasite-naive calves. The specificity of both the ELISA and IHA was very high, 99.2% and 99.6%, respectively. The sensitivity of the ELISA (100%) was significantly higher than that of the IHA (78.1%). In experimentally infected cattle, D. viviparus-specific antibodies were first detected with the ELISA between days 28 and 42 post-infection (p.i.), whereas the IHA only became positive between days 42 and 70. With the ELISA, antibody levels persisted until day 168 p.i. The IHA remained positive until the end of the experiment (day 196). None of the vaccinated animals were seropositive with the ELISA, whereas 25% of the calves were seropositive with the IHA. The seroprevalence of D. viviparus infections was determined in a field study with 467 sera from cattle of 64 herds; 227 (48.6%) of the animals were seropositive with the ELISA whereas only 38 (8.1%) scored positive with the IHA. To determine the precision of the ELISA, a total of five laboratories participated in trials, in which panels of strong positive, positive, and weak positive candidate sera were tested blind according to an international (International Standard ISO 5725, 1986) standard procedure. The repeatability and reproducibility of the ELISA were 0-16% and 14-26%, respectively. After these promising results it was decided to introduce this ELISA in 1995 as a routine test in all Animal Health Services in the Netherlands, replacing the IHA and faecal examinations for lungworm.
对一种用于检测实验感染和自然感染牛体内抗胎生网尾线虫抗体的酶联免疫吸附测定(ELISA)进行了评估,分析了其敏感性、特异性、血清转化时间、抗胎生网尾线虫反应的持久性以及精密度。将前三个参数与间接血凝试验(IHA)的相应参数进行了比较。在从实验感染了胎生网尾线虫、奥斯特他线虫、牛古柏线虫、瑞士细颈线虫、猪蛔虫或肝片吸虫纯分离株的犊牛以及未接触过寄生虫的犊牛采集的血清中,评估了两种检测方法的特异性和敏感性。ELISA和IHA的特异性都非常高,分别为99.2%和99.6%。ELISA的敏感性(100%)显著高于IHA(78.1%)。在实验感染的牛中,ELISA在感染后(p.i.)第28至42天首次检测到胎生网尾线虫特异性抗体,而IHA在第42至70天才呈阳性。使用ELISA时,抗体水平持续到感染后第168天。IHA在实验结束时(第196天)仍为阳性。ELISA检测中,没有一只接种疫苗的动物呈血清阳性,而IHA检测中有25%的犊牛呈血清阳性。在一项对64个牛群的467份血清进行的现场研究中,测定了胎生网尾线虫感染的血清阳性率;ELISA检测显示227只(48.6%)动物呈血清阳性,而IHA检测只有38只(8.1%)呈阳性。为了确定ELISA的精密度,共有五个实验室参与了试验,按照国际(国际标准ISO 5725,1986)标准程序对强阳性、阳性和弱阳性候选血清样本进行了盲法检测。ELISA的重复性和再现性分别为0 - 16%和14 - 26%。基于这些良好结果,1995年决定在荷兰所有动物卫生服务机构将这种ELISA作为常规检测方法引入,取代IHA和肺线虫粪便检查。