Marot-Leblond Agnes, Beucher Bertrand, David Sandrine, Nail-Billaud Sandrine, Robert Raymond
Groupe d'Etude des Interactions Hôte-Parasite, UPRES EA 3142, UFR des Sciences Pharmaceutiques et d'Ingénierie de la Santé, Angers, France.
J Clin Microbiol. 2006 Jan;44(1):138-42. doi: 10.1128/JCM.44.1.138-142.2006.
Cell components of the dimorphic pathogenic fungus Candida dubliniensis were used to prepare monoclonal antibodies (MAbs). One MAb, designated 12F7-F2, was shown by indirect immunofluorescence to be specific for a surface antigen of Candida dubliniensis yeast cells. No reactivity was observed with other fungal genera or with other Candida species, including Candida albicans, that share many phenotypic features with C. dubliniensis. The use of different chemical and physical treatments for cell component extraction suggested that the specific epitope probably resides on a protein moiety absent from C. albicans. However, we failed to identify the target protein by Western blotting, owing to its sensitivity to heat and sodium dodecyl sulfate. MAb 12F7-F2 was further used to develop a commercial latex agglutination test to identify C. dubliniensis colonies (Bichro-dubli Fumouze test; Fumouze Diagnostics). The test was validated on yeast strains previously identified by PCR and on fresh clinical isolates; these included 46 C. dubliniensis isolates, 45 C. albicans isolates, and other yeast species. The test had 100% sensitivity and specificity for C. dubliniensis isolated on Sabouraud dextrose, CHROMagar Candida, and CandiSelect media and 97.8% sensitivity for C. dubliniensis grown on Candida ID medium. The test is rapid (5 min) and easy to use and may be recommended for routine use in clinical microbiology laboratories and for epidemiological investigations.
利用双相致病性真菌都柏林念珠菌的细胞成分制备单克隆抗体(MAb)。其中一种单克隆抗体命名为12F7-F2,间接免疫荧光显示其对都柏林念珠菌酵母细胞的表面抗原有特异性。未观察到与其他真菌属或与都柏林念珠菌有许多表型特征相同的其他念珠菌物种(包括白色念珠菌)发生反应。对细胞成分提取采用不同的化学和物理处理方法表明,该特异性表位可能存在于白色念珠菌所没有的一种蛋白质部分上。然而,由于其对热和十二烷基硫酸钠敏感,我们未能通过蛋白质印迹法鉴定出靶蛋白。单克隆抗体12F7-F2进一步用于开发一种商业乳胶凝集试验,以鉴定都柏林念珠菌菌落(Bichro-dubli Fumouze试验;Fumouze诊断公司)。该试验在先前通过PCR鉴定的酵母菌株和新鲜临床分离株上进行了验证;这些分离株包括46株都柏林念珠菌、45株白色念珠菌和其他酵母菌种。该试验对在沙氏葡萄糖培养基、CHROMagar念珠菌培养基和CandiSelect培养基上分离的都柏林念珠菌的敏感性和特异性均为100%,对在念珠菌ID培养基上生长的都柏林念珠菌的敏感性为97.8%。该试验快速(5分钟)且易于使用,可推荐用于临床微生物实验室的常规使用和流行病学调查。