基于过氧化物酶信号扩增的重组抗原ELISA检测Q热特异性抗体

Hua-Wei Chen, Zhiwen Zhang, E. Glennon, W. Ching
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引用次数: 8

摘要

目前公认的Q热血清诊断方法是采用全细胞抗原的间接免疫荧光抗体测定法(IFA)。在本研究中,我们制备了27-kDa外膜蛋白(Com1)的重组抗原,该蛋白已被证实可被Q热患者血清识别。以IFA确认的血清样品,ELISA法评价重组Com1的性能。由于Q热患者IgG和IgM滴度较低,采用生物素化的抗人IgG或IgM加链亲和素- hrp聚合物进一步扩增标准ELISA信号。改良后的酶联免疫吸附试验(ELISA)能检测出88%(33人中29人)的驻伊拉克海军陆战队Q热患者血清。来自其他发热疾病患者的血清中只有不到5%(156例中有5例)与Com1反应。这些结果表明,使用Com1修饰的ELISA可能具有提高Q热特异性抗体检测的潜力。
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Detection of Q Fever Specific Antibodies Using Recombinant Antigen in ELISA with Peroxidase Based Signal Amplification
Currently, the accepted method for Q fever serodiagnosis is indirect immunofluorescent antibody assay (IFA) using the whole cell antigen. In this study, we prepared the recombinant antigen of the 27-kDa outer membrane protein (Com1) which has been shown to be recognized by Q fever patient sera. The performance of recombinant Com1 was evaluated in ELISA by IFA confirmed serum samples. Due to the low titers of IgG and IgM in Q fever patients, the standard ELISA signals were further amplified by using biotinylated anti-human IgG or IgM plus streptavidin-HRP polymer. The modified ELISA can detect 88% (29 out of 33) of Q fever patient sera collected from Marines deployed to Iraq. Less than 5% (5 out of 156) of the sera from patients with other febrile diseases reacted with the Com1. These results suggest that the modified ELISA using Com1 may have the potential to improve the detection of Q fever specific antibodies.
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