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酶联免疫吸附测定法和免疫印迹法在猪痢疾密螺旋体抗体检测中的应用及评估

Application and evaluation of enzyme-linked immunosorbent assay and immunoblotting for detection of antibodies to Treponema hyodysenteriae in swine.

作者信息

Smith S C, Barrett L M, Muir T, Christopher W L, Coloe P J

机构信息

Department of Applied Biology, Royal Melbourne Institute of Technology, Victoria, Australia.

出版信息

Epidemiol Infect. 1991 Oct;107(2):285-96. doi: 10.1017/s0950268800048937.

Abstract

An enzyme-linked immunoassay (ELISA) has been developed to detect serum Immunoglobulin antibodies G and M to Treponema hyodysenteriae in vaccinated, experimentally infected and naturally infected swine. Naturally infected swine gave ELISA titres that were similar to experimentally infected swine, but were significantly less than the titres of vaccinated swine. When serum from naturally infected swine was used to probe nitrocellulose blots of sodium dodecyl sulphate-polyacrylamide gel electrophoresed whole cell proteins of T. hyodysenteriae, the immunoblotting patterns showed IgG antibodies were produced against many T. hyodysenteriae protein antigens and against lipopolysaccharide (LPS). The IgG antibodies directed against LPS were serotype-specific for that LPS and could be used to identify the serotype involved in the T. hyodysenteriae infection in that herd. IgM immunoblots also reacted with the many protein antigens but were less specific for LPS antigen, with a substantial degree of cross-reaction between the LPS of all serotypes. The data demonstrate that a microplate enzyme-linked immunosorbent assay, coupled with immunoblotting, is a very specific and sensitive test for detection of antibody to Treponema hyodysenteriae in swine.

摘要

已开发出一种酶联免疫吸附测定(ELISA),用于检测接种疫苗、实验感染和自然感染猪血清中针对猪痢疾密螺旋体的免疫球蛋白G和M抗体。自然感染猪的ELISA滴度与实验感染猪相似,但明显低于接种疫苗猪的滴度。当用自然感染猪的血清探测经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳的猪痢疾密螺旋体全细胞蛋白的硝酸纤维素印迹时,免疫印迹模式显示产生了针对多种猪痢疾密螺旋体蛋白抗原和脂多糖(LPS)的IgG抗体。针对LPS的IgG抗体对该LPS具有血清型特异性,可用于鉴定该猪群中猪痢疾密螺旋体感染所涉及的血清型。IgM免疫印迹也与多种蛋白抗原发生反应,但对LPS抗原的特异性较低,所有血清型的LPS之间存在相当程度的交叉反应。数据表明,微孔板酶联免疫吸附测定结合免疫印迹是检测猪血清中猪痢疾密螺旋体抗体的一种非常特异和灵敏的试验。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92c1/2272052/159b34ba9cfa/epidinfect00029-0054-a.jpg

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