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一种利用与gra2蛋白的内在无序结构域相连的重组SAG1抗原N端一半进行弓形虫病快速诊断的检测方法。

A rapid diagnostic test for toxoplasmosis using recombinant antigenic N-terminal half of SAG1 linked with intrinsically unstructured domain of gra2 protein.

作者信息

Song Kyoung Ju, Yang Zhaoshou, Chong Chom-Kyu, Kim Jin-Soo, Lee Kyung Chan, Kim Tong-Soo, Nam Ho-Woo

机构信息

Department of Parasitology, Catholic Institute of Parasitic Disease, College of Medicine, Catholic University of Korea, Seoul 137-701, Korea.

出版信息

Korean J Parasitol. 2013 Oct;51(5):503-10. doi: 10.3347/kjp.2013.51.5.503. Epub 2013 Oct 31.

Abstract

Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis.

摘要

刚地弓形虫是一种顶复门寄生虫,宿主范围广泛,包括人类在内的大多数温血哺乳动物都能被其感染。全球有三分之一的人口已被感染,感染后可能导致先天性缺陷、流产和新生儿并发症。在此,我们开发了一种用于检测刚地弓形虫感染的快速诊断测试(RDT)。主要表面抗原(SAG1)的抗原性N端一半与致密颗粒蛋白2(GRA2)的内在无序结构域(IUD)相连。重组GST-GRA2-SAG1A蛋白成功表达并纯化,分子量为51 kDa。此外,rGST-GRA2-SAG1A蛋白的抗原性和溶解性显著提高。通过与以刚地弓形虫全细胞裂解物为抗原的ELISA结果进行比较,负载GST-GRA2-SAG1A的RDT(TgRDT)的总体特异性和敏感性估计分别为100%和97.1%。用乌干达人血清进行现场试验的TgRDT显示,弓形虫抗体血清阳性率为31.9%。TgRDT被证明是一种快速、易于使用且准确性高的试剂盒,将是一种适合用于弓形虫病的血清学诊断工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab5/3857496/672dd8abd735/kjp-51-503-g001.jpg

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