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GRA2 and ROP1 Recombinant Antigens as Potential Markers for Detection of Toxoplasma gondii-Specific Immunoglobulin G in Humans with Acute Toxoplasmosis

机译:GRA2和ROP1重组抗原作为检测弓形虫人急性弓形虫病中弓形虫特异性免疫球蛋白G的潜在标志物

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A goal of the current study was to evaluate serological applications of Toxoplasma gondii GRA2 and rhoptry protein 1 (ROP1) antigens. Soluble recombinant GRA2 and ROP1 antigens as fusion proteins containing six histidyl residues at the N and C terminals were obtained using an Escherichia coli expression system. Purification by one-step metal affinity chromatography allowed recovery of milligram amounts of pure recombinant proteins per liter of culture. The usefulness of these antigens for diagnosis of human infections was tested on 167 serum samples obtained during routine diagnostic tests. A panel of 37 serum samples from patients with acute toxoplasmosis was compared to a panel of 90 serum samples from individuals with past infection. The results indicated that both GRA2 and ROP1 recombinant antigens detected antibodies more frequently in samples from individuals with acute infections (100% and 94.6%, respectively) than in samples from individuals with chronic infections (22.5% and 15.5%, respectively). These results suggest that immunoglobulin G antibodies against GRA2 and ROP1 antigens are produced during the acute stage of toxoplasmosis but are uncommon in the chronic phase of the infection. Hence, these recombinant proteins can be used as specific molecular markers to differentiate between acute and chronic infections.
机译:本研究的目的是评估弓形虫GRA2和rhoptry蛋白1(ROP1)抗原的血清学应用。利用大肠杆菌表达系统,获得了可溶性重组GRA2和ROP1抗原,其融合蛋白在N和C末端含有六个组氨酸残基。通过一步金属亲和色谱纯化,每升培养物可回收毫克量的纯重组蛋白。在常规诊断测试中获得的167个血清样本中测试了这些抗原对人类感染诊断的有用性。将一组来自急性弓形虫病患者的37个血清样品与一组来自过去感染个体的90个血清样品进行比较。结果表明,GRA2和ROP1重组抗原在患有急性感染的个体(分别为100%和94.6%)的样品中检测抗体的频率比在患有慢性感染的个体(分别为22.5%和15.5%)的样品中检出抗体的频率更高。这些结果表明,在弓形虫病的急性期会产生针对GRA2和ROP1抗原的免疫球蛋白G抗体,但在感染的慢性期却很少见。因此,这些重组蛋白可用作特异性分子标记,以区分急性和慢性感染。

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