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展示白色念珠菌分泌天冬氨酸蛋白酶表位的杂交噬菌体的制备及其在播散性念珠菌病诊断中的应用。

Production of hybrid phage displaying secreted aspartyl proteinase epitope of Candida albicans and its application for the diagnosis of disseminated candidiasis.

作者信息

Yang Qiong, Su Quan-Ping, Wang Gui-Yun, Wen De-Zhong, Zhang Yan-Hui, Bao Hui-Zheng, Wang Li

机构信息

Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, Changchun City, Jilin Province, People's Republic of China.

出版信息

Mycoses. 2007 May;50(3):165-71. doi: 10.1111/j.1439-0507.2006.01349.x.

Abstract

The secreted aspartyl proteinases (Saps) of Candida albicans have been implicated as immunodominant antigens and virulence factors associated with adherence and tissue invasion. A hybrid phage displaying the Sap epitope VKYTS was constructed by cloning the corresponding DNA fragments into the pfd88 vector. Similar to native Sap, the phage-displayed epitope showed reactivity to sera from mice and patients with systemic C. albicans infection but not from those with oropharyngeal candidiasis and healthy individuals on Western blot. Furthermore, a new enzyme-linked immunosorbent assay was developed to detect the anti-Sap antibody with hybrid phage displaying Sap epitope VKYTS that can be recognised by anti-Sap antibodies. Sequential sera were tested from patients and mice with systemic candidiasis and oropharyngeal candidiasis, and serum samples from healthy individuals were also included. The sensitivity and specificity were 77% and 88.3% for experimental mice, respectively. These values reached 60% and 85%, respectively, for human patients. These data indicate this phage-displayed epitope as an effective and less expensive reagent would be a valuable probe for the detection of specific Sap antibody in the sera of patients and mice with systemic C. albicans infection.

摘要

白色念珠菌分泌的天冬氨酸蛋白酶(Saps)被认为是免疫显性抗原和与黏附及组织侵袭相关的毒力因子。通过将相应的DNA片段克隆到pfd88载体中构建了展示Sap表位VKYTS的杂交噬菌体。与天然Sap相似,噬菌体展示的表位在蛋白质印迹法中对来自患有全身性白色念珠菌感染的小鼠和患者的血清有反应,但对患有口咽念珠菌病的患者和健康个体的血清无反应。此外,开发了一种新的酶联免疫吸附测定法,以检测具有可被抗Sap抗体识别的展示Sap表位VKYTS的杂交噬菌体的抗Sap抗体。检测了全身性念珠菌病和口咽念珠菌病患者及小鼠的连续血清,还包括健康个体的血清样本。实验小鼠的敏感性和特异性分别为77%和88.3%。人类患者的这些值分别达到60%和85%。这些数据表明,这种噬菌体展示的表位作为一种有效且成本较低的试剂,将是检测全身性白色念珠菌感染患者和小鼠血清中特异性Sap抗体的有价值探针。

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