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基于 Vitek MS 系统使用和核糖体 DNA 测序的酵母鉴定算法:中国侵袭性真菌监测计划的参考检测建议。

Yeast identification algorithm based on use of the Vitek MS system selectively supplemented with ribosomal DNA sequencing: proposal of a reference assay for invasive fungal surveillance programs in China.

机构信息

Department of Clinical Laboratory, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, China.

出版信息

J Clin Microbiol. 2014 Feb;52(2):572-7. doi: 10.1128/JCM.02543-13. Epub 2013 Dec 11.

DOI:10.1128/JCM.02543-13
PMID:24478490
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3911352/
Abstract

Sequence analysis of the internal transcribed spacer (ITS) region was employed as the gold standard method for yeast identification in the China Hospital Invasive Fungal Surveillance Net (CHIF-NET). It has subsequently been found that matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is potentially a more practical approach for this purpose. In the present study, the performance of the Vitek MS v2.0 system for the identification of yeast isolates collected from patients with invasive fungal infections in the 2011 CHIF-NET was evaluated. A total of 1,243 isolates representing 31 yeast species were analyzed, and the identification results by the Vitek MS v2.0 system were compared to those obtained by ITS sequence analysis. By the Vitek MS v2.0 system, 96.7% (n = 1,202) of the isolates were correctly assigned to the species level and 0.2% (n = 2) of the isolates were identified to the genus level, while 2.4% (n = 30) and 0.7% (n = 9) of the isolates were unidentified and misidentified, respectively. After retesting of the unidentified and misidentified strains, 97.3% (n = 1,209) of the isolates were correctly identified to the species level. Based on these results, a testing algorithm that combines the use of the Vitek MS system with selected supplementary ribosomal DNA (rDNA) sequencing was developed and validated for yeast identification purposes. By employing this algorithm, 99.7% (1,240/1,243) of the study isolates were accurately identified with the exception of two isolates of Candida fermentati and one isolate of Cryptococcus gattii. In conclusion, the proposed identification algorithm could be practically implemented in strategic programs of fungal infection surveillance.

摘要

采用内转录间隔区(ITS)序列分析作为中国医院侵袭性真菌监测网(CHIF-NET)中酵母鉴定的金标准方法。随后发现基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)对于此目的可能是一种更实用的方法。本研究评估了 Vitek MS v2.0 系统用于鉴定 2011 年 CHIF-NET 中侵袭性真菌感染患者分离的酵母的性能。共分析了代表 31 种酵母的 1243 株分离株,将 Vitek MS v2.0 系统的鉴定结果与 ITS 序列分析的结果进行比较。通过 Vitek MS v2.0 系统,96.7%(n=1202)的分离株被正确分配到种水平,0.2%(n=2)的分离株被鉴定到属水平,而 2.4%(n=30)和 0.7%(n=9)的分离株未鉴定和鉴定错误。对未鉴定和鉴定错误的菌株进行复测后,97.3%(n=1209)的分离株被正确鉴定到种水平。基于这些结果,开发并验证了一种结合使用 Vitek MS 系统和选定的核糖体 DNA(rDNA)测序的测试算法,用于酵母鉴定。通过使用该算法,除了两株 Candida fermentati 和一株 Cryptococcus gattii 分离株外,研究中 99.7%(1240/1243)的分离株被准确鉴定。总之,所提出的鉴定算法可在真菌感染监测的战略计划中实际实施。

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