Molecular cloning and characterization of a recombinant Histoplasma capsulatum antigen for antibody-based diagnosis of human histoplasmosis
Molecular cloning and characterization of a recombinant Histoplasma capsulatum antigen for antibody-based diagnosis of human histoplasmosis
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DOI：
10.1128/jcm.35.5.1071-1076.1997
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发表时间：
1997-05-01
影响因子：
9.4
通讯作者：
Weil, GJ
中科院分区：
文献类型：
--
作者：
Chandrashekar, R;Curtis, KC;Weil, GJ
真菌间的免疫交叉反应性阻碍了组织胞浆菌病特异性血清学诊断方法的发展。我们报告的分子克隆和表征的荚膜组织胞浆菌cDNA（GH 17）编码的抗原具有免疫诊断潜力。GH 17是一个810 bp的cDNA，编码211个氨基酸残基的蛋白质。GH 17序列与GenBank中的其它序列几乎没有显著的同源性。Southern印迹分析表明，GH 17仅在H.荚膜免疫印迹表明，GH 17的蛋白产物（表达为140-kDa的β-半乳糖苷酶融合蛋白）被18/18例组织胞浆菌病患者血清中的抗体识别，但不被其他真菌感染的患者或动物血清中的抗体识别。在H的免疫印迹中，针对该蛋白质产生的抗体与60-kDa的天然抗原结合。荚膜酵母抗原提取物。这些结果表明GH 17编码一个H.荚膜抗原，可用于诊断人类组织胞浆菌病。
Immunological cross-reactivity among fungi has hampered the development of specific serodiagnostic assays for histoplasmosis. We report the molecular cloning and characterization of a Histoplasma capsulatum cDNA (GH17) that encodes an antigen with immunodiagnostic potential. GH17 is an 810-bp cDNA which encodes a protein of 211 amino acid residues. The GH17 sequence has almost no significant homology with other sequences in GenBank. Southern blot analysis suggests that GH17 is confined to a single location in the genomic DNA of H. capsulatum. Immunoblots indicated that the protein product of GH17 (expressed as a 140-kDa beta-galactosidase fusion protein) was recognized by antibodies in 18 of 18 sera from histoplasmosis patients, but not by antibodies in sera from patients or animals infected with other fungi, GH17 was expressed in a prokaryotic expression vector, pPRoEX-1, and recombinant protein was purified by preparative electrophoresis. Antibodies raised to this protein bound to a 60-kDa native antigen in immunoblots of H. capsulatum yeast antigen extract. These results suggest that GH17 encodes an H. capsulatum antigen that may be useful for the diagnosis of histoplasmosis in humans.