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在大肠杆菌中表达的刚地弓形虫重组棒状体抗原8的血清学诊断评估

Sero-diagnostic evaluation of Toxoplasma gondii recombinant Rhoptry antigen 8 expressed in E. coli.

作者信息

Sonaimuthu Parthasarathy, Fong Mun Yik, Kalyanasundaram Ramaswamy, Mahmud Rohela, Lau Yee Ling

机构信息

Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

出版信息

Parasit Vectors. 2014 Jul 1;7:297. doi: 10.1186/1756-3305-7-297.

Abstract

BACKGROUND

Toxoplasma gondii infects all warm-blooded animals, including humans. Early diagnosis and determining the infective stage are critical for effectively treating immunosuppressed individuals and pregnant women with toxoplasmosis. Among the rhoptry proteins of the parasite, Rhoptry protein 8 (ROP8), is known to be expressed during the early stages of T. gondii infection and is involved in parasitophorous vacuole formation. In this study, we have investigated the diagnostic efficacy of recombinant ROP8 (rROP8).

METHODS

The ROP8 gene was cloned into pCOLD I DNA vector and expressed as a soluble recombinant antigen in Escherichia coli. Expressed ROP8 protein was evaluated using western blot method.

RESULTS

Western blot analysis of purified rROP8 antigen using 200 T. gondii-infected human serum samples, as well as non-infected serum controls, allowed for the successful identification of toxoplasmosis-positives, yielding a 90% sensitivity and 94% specificity.

CONCLUSION

Our findings indicated that rROP8 antigen expressed in E. coli was able to detect toxoplasmosis in infected human serum with specificity and sensitivity suggesting that rROP8 antigen represents a valid diagnostic marker for toxoplasmosis.

摘要

背景

刚地弓形虫可感染包括人类在内的所有温血动物。早期诊断并确定感染阶段对于有效治疗弓形虫病免疫抑制个体和孕妇至关重要。在该寄生虫的棒状体蛋白中,棒状体蛋白8(ROP8)已知在刚地弓形虫感染早期表达,并参与寄生泡的形成。在本研究中,我们调查了重组ROP8(rROP8)的诊断效力。

方法

将ROP8基因克隆到pCOLD I DNA载体中,并在大肠杆菌中作为可溶性重组抗原表达。使用蛋白质印迹法评估表达的ROP8蛋白。

结果

使用200份刚地弓形虫感染的人血清样本以及未感染血清对照对纯化的rROP8抗原进行蛋白质印迹分析,成功鉴定出弓形虫病阳性样本,灵敏度为90%,特异性为94%。

结论

我们的研究结果表明,在大肠杆菌中表达的rROP8抗原能够特异性且灵敏地检测感染人血清中的弓形虫病,这表明rROP8抗原是弓形虫病的有效诊断标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b5d4/4098696/ea352a5d2ca0/1756-3305-7-297-1.jpg

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