Department of Virology, Parasitology and Immunology, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
Vet Parasitol. 2012 Oct 26;189(2-4):267-73. doi: 10.1016/j.vetpar.2012.04.024. Epub 2012 Apr 23.
The significant economical consequences of infections with Ascaris suum in pigs are already well documented. However, due to the subclinical nature of the disease and the lack of practical diagnostic means, ascariasis often remains undiagnosed. Here we describe the development and evaluation of a novel indirect ELISA using the purified A. suum haemoglobin (AsHb) molecule as an antigen. Initial validation using sera from 190 pigs experimentally infected twice a week with A. suum and Trichuris suis (25 and 5 eggs kg(-1)day(-1) respectively) demonstrated that the AsHb ELISA is able to detect long-term exposure to A. suum with a high sensitivity and specificity (99.5% and 100.0% respectively). Furthermore, this serological technique proved to be more sensitive than faecal examination on week 7 and 14 of the experiment (99.5% and 100% compared to 59.5% and 68.4% respectively). Cross-reactivity caused by T. suis infection was shown to be limited after analysing sera from pigs with an experimental T. suis mono-infection. Seroconversion was shown to occur from week 6 onwards in pigs receiving 100 A. suum eggs 5 times a week. Preliminary testing of the ELISA on six randomly selected farms confirmed the results obtained in the artificial infection trials, showing a higher sensitivity of the serologic method compared to faecal examination. Finally, the ELISA was used to investigate Ascaris infection rates on 101 conventional Flemish pig farms. The results showed that on 38.6% of the farms less than 20% of the tested samples were seropositive, while in 19.8% of the farms 80-100% of all pigs were seropositive. The results of this study suggest that the AsHb ELISA could provide pig farmers and veterinarians with an easier and more sensitive way to estimate the overall prevalence of A. suum on their farm.
猪感染蛔虫造成的显著经济后果已得到充分证实。然而,由于疾病的亚临床性质和缺乏实用的诊断手段,蛔虫病常常未被诊断。在此,我们描述了一种新型间接 ELISA 的开发和评估,该 ELISA 使用纯化的蛔虫血红蛋白(AsHb)分子作为抗原。使用 190 头每周两次用蛔虫和猪鞭虫(分别为 25 和 5 个卵/公斤体重/天)感染的猪的血清进行初步验证,结果表明 AsHb ELISA 能够检测到长期暴露于蛔虫,具有高灵敏度和特异性(分别为 99.5%和 100.0%)。此外,与实验第 7 周和第 14 周的粪便检查相比,该血清学技术被证明更为敏感(分别为 99.5%和 100%比 59.5%和 68.4%)。分析患有实验性猪鞭虫单感染的猪的血清表明,由猪鞭虫感染引起的交叉反应是有限的。每周接受 100 个蛔虫卵 5 次的猪从第 6 周开始显示出血清转化。对六个随机选择的农场进行的 ELISA 初步测试证实了人工感染试验的结果,表明与粪便检查相比,血清学方法的灵敏度更高。最后,该 ELISA 用于调查 101 个常规佛兰芒猪农场的蛔虫感染率。结果表明,在 38.6%的农场中,不到 20%的测试样本呈血清阳性,而在 19.8%的农场中,80-100%的猪呈血清阳性。本研究结果表明,AsHb ELISA 可以为养猪户和兽医提供一种更简单、更敏感的方法来估计其农场蛔虫的总体流行率。