柔嫩艾美耳球虫重组融合蛋白的制备及其在酶联免疫吸附测定中的诊断潜力评估

Production of a recombinant fusion protein of Sarcocystis tenella and evaluation of its diagnostic potential in an ELISA.

作者信息

Mertens C M, Tenter A M, Vietmeyer C, Ellis J T, Johnson A M

机构信息

Institut für Parasitologie, Tierärztliche Hochschule Hannover, Germany.

出版信息

Vet Parasitol. 1996 Oct 25;65(3-4):185-97. doi: 10.1016/s0304-4017(96)00971-5.

DOI:10.1016/s0304-4017(96)00971-5

PMID:8983145

Abstract

We have isolated a cDNA clone encoding an antigenic polypeptide of Sarcocystis tenella by screening a cystozoite-derived cDNA library in lambda gt11 with antibodies from sheep infected experimentally with S. tenella. This clone, termed STC29, was subcloned and expressed in the vector pGEX-3X as a soluble fusion protein with glutathione S-transferase having an apparent molecular mass of 46 kDa. Antibody raised against the recombinant fusion protein recognized a native polypeptide of 25 kDa in cystozoites of S. tenella. In an ELISA with sera from experimentally infected sheep, the recombinant STC29 antigen could differentiate infections with S. tenella from those with Sarcocystis arieticanis or Toxoplasma gondii. Hence, the research described here reports the identification of the first recombinant S. tenella antigen that may be useful for standardization of a serological test for the diagnosis of S.tenella infections in sheep.

摘要

我们通过用经实验感染柔嫩艾美耳球虫的绵羊产生的抗体筛选λgt11载体中的子孢子衍生cDNA文库，分离出了一个编码柔嫩艾美耳球虫抗原多肽的cDNA克隆。这个名为STC29的克隆被亚克隆并在载体pGEX - 3X中表达，作为与谷胱甘肽S - 转移酶的可溶性融合蛋白，其表观分子量为46 kDa。针对重组融合蛋白产生的抗体识别出柔嫩艾美耳球虫子孢子中一条25 kDa的天然多肽。在一项针对经实验感染绵羊血清的ELISA检测中，重组STC29抗原能够区分柔嫩艾美耳球虫感染与阿氏艾美耳球虫或刚地弓形虫感染。因此，此处描述的研究报告了首个重组柔嫩艾美耳球虫抗原的鉴定，该抗原可能有助于标准化用于诊断绵羊柔嫩艾美耳球虫感染的血清学检测。