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Serological diagnosis of pneumocystosis: production of a synthetic recombinant antigen for immunodetection of Pneumocystis jirovecii

机译:肺囊虫病的血清学诊断:合成重组抗原的生产,用于免疫检测吉氏肺孢子虫

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Diagnosis of Pneumocystis pneumonia (PcP) relies on the detection of P. jirovecii in respiratory specimens obtained by invasive techniques. Thus, the development of a serological test is urgently needed as it will allow the diagnosis of PcP using blood, an inexpensive and non-invasive specimen. This study aims to combine the production of a multi-epitope synthetic recombinant antigen (RSA) and an ELISA test for detection of anti-P. jirovecii antibodies, in order to develop a new approach for PcP diagnosis. The RSA was selected and designed based on the study of the immunogenicity of the carboxyl-terminal domain of the major surface glycoprotein. This antigen was purified and used as an antigenic tool in an ELISA technique for detection of Ig, IgG and IgM antibodies anti-P. jirovecii (patent-pending no. PT109078). Serum specimens from 88 patients previously categorized in distinct clinical subgroups and 17 blood donors, were analysed. The IgM anti-P. jirovecii levels were statistically increased in patients with PcP (p?=?0.001) and the ELISA IgM anti-P. jirovecii test presented a sensitivity of 100% and a specificity of 80.8%, when associated with the clinical diagnosis criteria. This innovative approach, provides good insights about what can be done in the future serum testing for PcP diagnosis.
机译:肺囊虫性肺炎(PcP)的诊断依赖于通过侵入性技术获得的呼吸道标本中的jirovecii疟原虫的检测。因此,迫切需要开发血清学检测试剂,因为它将允许使用血液(一种廉价且无创的标本)诊断PcP。这项研究的目的是将多表位合成重组抗原(RSA)的生产与用于检测抗P的ELISA试验结合起来。 jirovecii抗体,以开发一种新的PcP诊断方法。基于对主要表面糖蛋白的羧基末端结构域的免疫原性的研究,选择并设计了RSA。纯化该抗原,并将其用作ELISA技术中的抗原性工具,用于检测抗P的Ig,IgG和IgM抗体。 jirovecii(专利申请号PT109078)。分析了先前分类为不同临床亚组的88位患者的血清标本和17位献血者的血清标本。 IgM抗P。 PcP和ELISA IgM抗-P患者的jirovecii水平在统计学上增加(p≥0.001)。当与临床诊断标准相关联时,jirovecii试验的敏感性为100%,特异性为80.8%。这种创新的方法为将来在进行PcP诊断的血清检测中可以做什么提供了很好的见识。

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