Lischewski A, Kretschmar M, Hof H, Amann R, Hacker J, Morschhäuser J
Zentrum für Infektionsforschung, Universität Würzburg, Germany.
J Clin Microbiol. 1997 Nov;35(11):2943-8. doi: 10.1128/jcm.35.11.2943-2948.1997.
Two 18S rRNA-targeted oligonucleotide probes specific for Candida albicans and Candida parapsilosis were used to detect and identify by fluorescent in situ hybridization these medically important Candida species in deep organs of mice after experimental systemic infection. The C. albicans-specific probe detected fungal cells in kidney, spleen, and brain sections of a mouse infected with C. albicans but not in a mouse infected with the closely related species C. parapsilosis. Conversely, the C. parapsilosis-specific probe detected fungal cells in the deep organs of a mouse infected with C. parapsilosis but not in the deep organs of a C. albicans-infected mouse. In addition, the C. albicans-specific probe was used to detect this species in human blood spiked with yeast cells by a lysis-filtration assay and subsequent fluorescent in situ hybridization. By this assay, as few as three yeast cells per 0.5 ml of blood were consistently detected. Our results demonstrate that fluorescent in situ hybridization with species-specific rRNA-targeted oligonucleotide probes provides a novel, culture-independent method for the sensitive detection and identification of Candida species in clinically relevant material.
使用两种针对白色念珠菌和近平滑念珠菌的18S rRNA靶向寡核苷酸探针，通过荧光原位杂交来检测和鉴定实验性全身感染后小鼠深部器官中这些具有医学重要性的念珠菌属物种。白色念珠菌特异性探针在感染白色念珠菌的小鼠的肾脏、脾脏和脑切片中检测到真菌细胞，但在感染密切相关物种近平滑念珠菌的小鼠中未检测到。相反，近平滑念珠菌特异性探针在感染近平滑念珠菌的小鼠的深部器官中检测到真菌细胞，但在感染白色念珠菌的小鼠的深部器官中未检测到。此外，白色念珠菌特异性探针通过裂解过滤测定法和随后的荧光原位杂交用于检测掺入酵母细胞的人血中的该物种。通过该测定法，每0.5 ml血液中低至三个酵母细胞始终能够被检测到。我们的结果表明，用物种特异性rRNA靶向寡核苷酸探针进行荧光原位杂交提供了一种新颖的、无需培养的方法，用于在临床相关材料中灵敏地检测和鉴定念珠菌属物种。
