Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands.
Vet Microbiol. 2010 May 19;142(3-4):401-7. doi: 10.1016/j.vetmic.2009.11.003. Epub 2009 Nov 13.
The aim of this study is the development and evaluation of a serodiagnostic assay for Mycobacterium avium (MA). After screening MA lipid fractions in an ELISA format, a polar lipid fraction was selected as antigen because of its superior recognition by serum antibodies in experimentally infected pigs. The resulting MA-ELISA was evaluated as an alternative for detection of MA infection by traditional pathological examination of pig lymph nodes for granulomatous lesions by meat inspectors. By comparing with bacteriological examination, the MA-ELISA showed significantly better sensitivity (69%) as compared to pathological examination (31%) in experimentally infected pigs. The MA-ELISA also appeared significantly more specific in a set of serum samples from MA negative pigs: only 1 out of these 153 serum samples reacted positive, whereas 99 (65%) of these had displayed false positive results by detection of lymph nodes lesions that appeared not to be associated with MA (Komijn et al., 2007). The MA-ELISA was subsequently evaluated using serum samples from two farms with pigs known to be infected with MA. Bacteriological examination of the sub-maxillary and mesenteric lymph nodes showed that 56% (103/184) and 35% (41/117) of the pigs, respectively were positive for MA in these farms. In the first farm, 16% (29/184) of the pigs tested positive in MA-ELISA and 31% (57/184) by pathological examination. On the contrary, in the second farm, more pigs tested positive 17% (15/117) in MA-ELISA with 8% (9/117) positivity by pathological examination. Taking the results on both farms together, the sensitivity of the MA-ELISA was 14% and the specificity 83%, whereas the sensitivity of the pathological examination was 31% and the specificity 86%. For practical reasons use of a serological test as the MA-ELISA may be preferred over pathological or bacteriological examinations. Our studies in experimentally infected and negative "field" sera indicate that the MA-ELISA is significantly more specific and more sensitive than detection by classical pathological examination. However, the studies in two MA infected farms show a variable picture with pathological examination overall performing better. Study in a wider range of "positive" farms will be needed to provide a more comprehensive view of the quality of both tests for detection of MA in infected farms. At the same time further optimization of MA-ELISA with use of lipid antigens from a broader range of serotypes may improve its performance in the face of infections with different MA serotypes.
本研究的目的是开发和评估一种用于鸟分枝杆菌(MA)的血清诊断检测方法。在用 ELISA 筛选 MA 脂质部分后,选择了极性脂质部分作为抗原,因为它在实验感染的猪血清抗体中具有更好的识别能力。由此产生的 MA-ELISA 被评估为替代传统病理学检查通过肉检员对猪淋巴结肉芽肿病变的检测,以检测 MA 感染。通过与细菌学检查相比,MA-ELISA 在实验感染猪中的敏感性(69%)明显优于病理学检查(31%)。在一组来自 MA 阴性猪的血清样本中,MA-ELISA 也表现出明显更高的特异性:这些 153 个血清样本中只有 1 个呈阳性反应,而 99 个(65%)的血清样本通过检测似乎与 MA 无关的淋巴结病变呈假阳性结果(Komijn 等人,2007 年)。随后,使用来自已知感染 MA 的两个农场的血清样本对 MA-ELISA 进行了评估。颌下和肠系膜淋巴结的细菌学检查显示,在这些农场中,分别有 56%(103/184)和 35%(41/117)的猪 MA 呈阳性。在第一个农场,16%(29/184)的猪在 MA-ELISA 中呈阳性,31%(57/184)的猪在病理学检查中呈阳性。相反,在第二个农场,更多的猪在 MA-ELISA 中呈阳性 17%(15/117),而在病理学检查中呈阳性 8%(9/117)。将两个农场的结果结合起来,MA-ELISA 的敏感性为 14%,特异性为 83%,而病理学检查的敏感性为 31%,特异性为 86%。出于实际原因,使用血清学检测方法(如 MA-ELISA)可能优于病理学或细菌学检测。我们对实验感染和“现场”阴性血清的研究表明,MA-ELISA 比经典病理学检查具有更高的特异性和敏感性。然而,在两个 MA 感染农场的研究中,情况各不相同,总体而言,病理学检查的效果更好。在更广泛的“阳性”农场中进行研究,将提供对两种检测方法在感染农场中检测 MA 的质量的更全面了解。同时,使用更广泛的血清型脂质抗原进一步优化 MA-ELISA 可能会提高其在不同 MA 血清型感染下的性能。