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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Lipopolysaccharide-Based Enzyme-Linked Immunosorbent Assay for Experimental Use in Detection of Antibodies to Lawsonia intracellularis in Pigs
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Lipopolysaccharide-Based Enzyme-Linked Immunosorbent Assay for Experimental Use in Detection of Antibodies to Lawsonia intracellularis in Pigs

机译：基于脂多糖的酶联免疫吸附试验，用于检测猪胞内劳森菌的抗体

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An enzyme-linked immunosorbent assay (ELISA) for Lawsonia intracellularis was developed and compared with a whole-cell antigen-based immunofluorescence antibody test (IFAT). The antigen-containing lipopolysaccharide (LPS) was derived from Percoll gradient purified cultures of L. intracellularis by using a modification of the Westphal hot phenol procedure. The antigen was bound directly to polystyrene 96-well microtiter plates, and the assay was performed in an indirect ELISA format. Specificity and sensitivity values based on 80 known positive and 80 known negative serum samples from controlled experimental trials were 93.7% and 88.7%, respectively. Serological results from a controlled L. intracellularis challenge exposure study confirmed the high specificity and sensitivity of this assay (100% and 99.5%, respectively). Comparisons between the LPS ELISA and the IFAT in detecting anti-Lawsonia antibodies in this controlled study revealed significantly more LPS ELISA-positive pigs than IFAT-positive pigs on days 21, 28, 35, and 42 (P = 0.003, 0.030, 0.002, and 0.006, respectively). This indirect ELISA (LPS ELISA) test is an improved method of detecting antibodies in pigs soon after exposure to L. intracellularis, regardless of isolate type (vaccine or wild type) in experimental studies. The LPS ELISA may be used as a tool to support future research trials on vaccine efficacy and to further understand the immune response induced by L. intracellularis.
机译：酶联免疫吸附法（ELISA）的细胞内Lawsonia 进行了开发，并与基于全细胞抗原的免疫荧光抗体测试（IFAT）进行了比较。含抗原的脂多糖（LPS）源自Percoll梯度纯化的Lem培养物。韦斯特法热酚法的改进方法抗原直接与聚苯乙烯96孔微量滴定板结合，并以间接ELISA形式进行测定。根据来自对照实验的80个已知阳性和80个已知阴性血清样本的特异性和敏感性值分别为93.7％和88.7％。受控L血清学结果。细胞内挑战暴露研究证实了该检测方法的高特异性和敏感性（分别为100％和99.5％）。在这项对照研究中，LPS ELISA和IFAT在检测抗 Lawsonia 抗体方面的比较显示，在21、28、35和42天，LPS ELISA阳性猪明显多于IFAT阳性猪（< em> P 分别为0.003、0.030、0.002和0.006）。间接ELISA（LPS ELISA）测试是一种改进的方法，可在暴露于 L后立即检测猪中的抗体。实验研究中，不论分离株是疫苗型还是野生型，都可以检测到LPS ELISA可以用作支持疫苗功效的未来研究试验和进一步了解 L诱导的免疫反应的工具。细胞内。

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