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首页> 外文期刊>Journal of Clinical Microbiology >Evaluation of Fluorescent Capillary Electrophoresis for Rapid Identification of Candida Fungal Infections
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Evaluation of Fluorescent Capillary Electrophoresis for Rapid Identification of Candida Fungal Infections

机译：快速鉴定假丝酵母真菌感染的荧光毛细管电泳评估。

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Early diagnosis of fungal infection is critical for initiating antifungal therapy and reducing the high mortality rate in immunocompromised patients. In this study, we focused on rapid and sensitive identification of clinically important Candida species, utilizing the variability in the length of the ITS2 rRNA gene and fluorescent capillary electrophoresis (f-ITS2-PCR-CE). The method was developed and optimized on 29 various Candida reference strains from which 26 Candida species were clearly identified, while Candida guilliermondii, C. fermentati, and C. carpophila, which are closely related, could not be distinguished. The method was subsequently validated on 143 blinded monofungal clinical isolates (comprising 26 species) and was able to identify 88% of species unambiguously. This indicated a higher resolution power than the classical phenotypic approach which correctly identified 73%. Finally, the culture-independent potential of this technique was addressed by the analysis of 55 retrospective DNA samples extracted directly from clinical material. The method showed 100% sensitivity and specificity compared to those of the combined results of cultivation and panfungal PCR followed by sequencing used as a gold standard. In conclusion, this newly developed f-ITS2-PCR-CE analytical approach was shown to be a fast, sensitive, and highly reproducible tool for both culture-dependent and culture-independent identification of clinically important Candida strains, including species of the “psilosis” complex.
机译：真菌感染的早期诊断对于启动抗真菌治疗和降低免疫受损患者的高死亡率至关重要。在这项研究中，我们致力于利用 ITS2 rRNA基因长度的变异性和荧光毛细管电泳（f- ITS2 -PCR-CE）。该方法在29种不同的念珠菌参考菌株上进行了开发和优化，从中明确鉴定出26种念珠菌，而亲缘关系较近的念珠菌C. fermentati和C. carpophila则无法区分。该方法随后在143种盲单真菌临床分离株（包含26种）上得到了验证，并且能够明确鉴定88％的物种。这表明比传统的表型方法具有更高的分辨率，后者可以正确地识别出73％。最后，通过分析直接从临床材料中提取的55个回顾性DNA样品，分析了该技术的文化独立潜力。该方法与培养和Panfungal PCR结合后的测序结果（作为金标准）的结合结果相比显示出100％的敏感性和特异性。总之，这种新开发的f- ITS2 -PCR-CE分析方法被证明是一种快速，灵敏且可重复性高的工具，可用于对临床上重要的念珠菌菌株进行培养依赖性和非培养性鉴定，包括“矽肺”综合症的种类。

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