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评估荧光原位杂交(FISH)在鉴定白色念珠菌方面的应用,并与三种表型方法进行比较。

Evaluation of fluorescence in situ hybridisation (FISH) for the identification of Candida albicans in comparison with three phenotypic methods.

机构信息

Institute of Medical Microbiology and Hygiene, University of Ulm, Germany.

出版信息

Mycoses. 2012 May;55(3):e114-23. doi: 10.1111/j.1439-0507.2011.02154.x. Epub 2011 Nov 29.

DOI:10.1111/j.1439-0507.2011.02154.x
PMID:22126597
Abstract

Severe Candida infections are increasing and are associated with considerable morbidity and mortality. Rapid and accurate differentiation of Candida albicans from non-C. albicans species is essential for therapeutic decisions. We therefore developed a fluorescence in situ hybridisation (FISH) assay comprising previously described probes and a newly designed specific C. albicans probe/competitor probe combination. The FISH probes were first evaluated using 99 selected fungal strains covering 31 species, and a specificity between 96% and 100% and a sensitivity of 100%. The FISH assay was then applied to 110 clinical isolates in parallel with API32C, the chromogenic Candida ID agar, and determination of filamentous colony morphology. All tests produced highly reliable results. However, the Candida ID agar misidentified Candida dubliniensis as C. albicans. Determination of filamentous colony morphology allowed 100% reliable identification of C. albicans, but took 48 h. FISH allowed identification of clinical C. albicans isolates within 3 h with a sensitivity and specificity of 100%. FISH was additionally applied to 48 blood cultures showing yeasts in the Gram stain and correctly identified all 33 cases of C. albicans.

摘要

严重的念珠菌感染正在增加,并与相当大的发病率和死亡率相关。快速准确地区分白色念珠菌和非白色念珠菌物种对于治疗决策至关重要。因此,我们开发了一种荧光原位杂交(FISH)检测方法,包括以前描述的探针和新设计的特异性白色念珠菌探针/竞争探针组合。FISH 探针首先使用 99 株选定的真菌菌株进行评估,这些菌株涵盖 31 个物种,特异性为 96%至 100%,灵敏度为 100%。然后,将 FISH 检测方法与 API32C、显色念珠菌 ID 琼脂和丝状菌落形态的确定一起应用于 110 株临床分离株。所有测试都产生了高度可靠的结果。然而,念珠菌 ID 琼脂错误地将杜波依斯念珠菌鉴定为白色念珠菌。丝状菌落形态的确定可以 100%可靠地鉴定白色念珠菌,但需要 48 小时。FISH 可以在 3 小时内鉴定临床白色念珠菌分离株,其灵敏度和特异性均为 100%。FISH 还应用于 48 份显示革兰氏染色中有酵母的血培养物,正确鉴定了所有 33 例白色念珠菌感染。

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