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牛血清 22.6 抗原的分子克隆、鉴定及诊断性能。

Molecular cloning, characterization and diagnostic performance of the Schistosoma bovis 22.6 antigen.

机构信息

Parasitology Laboratory, Instituto de Recursos Naturales y Agrobiología de Salamanca (IRNASA, CSIC), Cordel de Merinas, 40-52, 37008 Salamanca, Spain.

出版信息

Vet Parasitol. 2012 Dec 21;190(3-4):530-40. doi: 10.1016/j.vetpar.2012.06.023. Epub 2012 Jun 26.

Abstract

Animal schistosomiasis caused by Schistosoma bovis is a veterinary problem in many areas of the world. It affects a large number of animals and causes important economic losses in livestock production. The 22.6 kDa antigen is a tegumental protein of unknown function, restricted to schistosomes. In S. bovis it has been identified in the tegument and in an excretion-secretion extract, consisting of several, non-glycosylated isoforms that are recognised by the sera of animals infected with S. bovis. The aims of the present work were to clone, sequence, express and characterize at molecular level the S. bovis 22.6 antigen (Sb22.6), as well as to assess the usefulness of the corresponding recombinant protein as a diagnostic antigen in ELISA tests for the detection of free-range cattle farms infested with S. bovis. Immunolocalization studies revealed that Sb22.6 is expressed in the tegument and some internal tissues of the adult worms, but it is not exposed on the surface of the adult worms and schistosomula. The reactivity of the recombinant Sb22.6 (rSb22.6) in ELISA against antibodies in sera from S. bovis experimentally infected hamsters and sera from free-range cattle from a S. bovis endemic area showed that the recombinant protein and the soluble extract of adult worms (SbC) exhibited a similar diagnostic performance. In addition, rSb22.6 did not show cross-reactions with antibodies against Fasciola hepatica, also a frequent trematode parasite in cattle. The rSb22.6 antigen can be readily produced in large amounts and in a highly reproducible fashion, avoiding the types of problem that arise upon using crude extracts such as the SbC. In conclusion, this protein represents a promising epidemiological tool for the surveillance of S. bovis and may help to implement control measures in the areas and farms were the parasite is present.

摘要

牛血吸虫病是一种由牛血吸虫引起的动物血吸虫病,在世界许多地区都是一个兽医问题。它影响了大量的动物,并在畜牧业生产中造成了重要的经济损失。22.6 kDa 抗原是一种未知功能的表皮蛋白,仅限于血吸虫。在 S. bovis 中,它已在表皮和排泄-分泌提取物中被鉴定出来,该提取物由几种非糖基化的同工型组成,这些同工型被感染 S. bovis 的动物血清所识别。本研究的目的是克隆、测序、表达和在分子水平上表征 S. bovis 22.6 抗原(Sb22.6),并评估相应的重组蛋白作为 ELISA 检测用于检测受 S. bovis 感染的自由放养牛场的有用性。免疫定位研究表明,Sb22.6 在成虫的表皮和一些内部组织中表达,但不在成虫和尾蚴的表面暴露。重组 Sb22.6(rSb22.6)在 ELISA 中对 S. bovis 实验感染仓鼠血清和 S. bovis 流行地区自由放养牛血清中的抗体的反应性表明,重组蛋白和成虫可溶性提取物(SbC)表现出相似的诊断性能。此外,rSb22.6 与针对 Fasciola hepatica 的抗体没有交叉反应,Fasciola hepatica 也是牛中一种常见的吸虫寄生虫。rSb22.6 抗原可以大量且高度重现地生产,避免了使用粗提物(如 SbC)时出现的问题。总之,该蛋白代表了一种有前途的监测 S. bovis 的流行病学工具,并可能有助于在寄生虫存在的地区和农场实施控制措施。

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