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建立和验证用于通过 MALDI-TOF MS 对酵母进行鉴定的扩展数据库,以应用于阿根廷。

Development and validation of an extended database for yeast identification by MALDI-TOF MS in Argentina.

机构信息

Departamento Micología, Instituto Nacional de Enfermedades Infecciosas "Dr. Carlos G. Malbrán," Ciudad Autónoma de Buenos Aires.

División Micología, Laboratorio de Salud Pública de Tucumán, Tucumán.

出版信息

Med Mycol. 2019 Feb 1;57(2):215-225. doi: 10.1093/mmy/myy021.

DOI:10.1093/mmy/myy021
PMID:29762695
Abstract

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) has revolutionized the identification of microorganisms in clinical laboratories because it is rapid, relatively simple to use, accurate, and can be used for a wide number of microorganisms. Several studies have demonstrated the utility of this technique in the identification of yeasts; however, its performance is usually improved by the extension of the database. Here we developed an in-house database of 143 strains belonging to 42 yeast species in the MALDI Biotyper platform, and we validated the extended database with 388 regional strains and 15 reference strains belonging to 55 yeast species. We also performed an intra- and interlaboratory study to assess reproducibility and analyzed the use of the cutoff values of 1.700 and 2.000 to correctly identify at species level. The creation of an in-house database that extended the manufacturer's database was successful in view of no incorrect identification was introduced. The best performance was observed by using the extended database and a cutoff value of 1.700 with a sensitivity of .94 and specificity of .96. A reproducibility study showed utility to detect deviations and could be used for external quality control. The extended database was able to differentiate closely related species and it has potential in distinguishing the molecular genotypes of Cryptococcus neoformans and Cryptococcus gattii.

摘要

基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术在临床实验室中极大地改变了微生物的鉴定方式,因为它快速、相对简单易用、准确,并且可以用于大量的微生物。多项研究已经证明了该技术在鉴定酵母方面的实用性;然而,通过扩展数据库通常可以提高其性能。在这里,我们在 MALDI Biotyper 平台上开发了一个包含 143 株属于 42 种酵母的内部数据库,并使用 388 株地区菌株和 15 株属于 55 种酵母的参考菌株验证了扩展数据库。我们还进行了一项室内和实验室间研究,以评估可重复性,并分析使用 1.700 和 2.000 的截止值来正确鉴定物种水平的效果。考虑到没有引入错误鉴定,创建扩展制造商数据库的内部数据库是成功的。使用扩展数据库和 1.700 的截止值观察到最佳性能,灵敏度为.94,特异性为.96。重复性研究表明该方法可用于检测偏差,可用于外部质量控制。扩展数据库能够区分密切相关的物种,并且有可能区分新型隐球菌和格特隐球菌的分子基因型。

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