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使用省时直接转移方案的两种基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)系统对从血流感染和脑脊液中分离出的酵母进行鉴定的性能

Performance of two MALDI-TOF MS systems for the identification of yeasts isolated from bloodstream infections and cerebrospinal fluids using a time-saving direct transfer protocol.

作者信息

Hamprecht Axel, Christ Sara, Oestreicher Tanja, Plum Georg, Kempf Volkhard A J, Göttig Stephan

机构信息

Institute for Medical Microbiology, Immunology and Hygiene, University Hospital of Cologne, Cologne, Germany,

出版信息

Med Microbiol Immunol. 2014 Apr;203(2):93-9. doi: 10.1007/s00430-013-0319-9. Epub 2013 Dec 6.

DOI:10.1007/s00430-013-0319-9
PMID:24310420
Abstract

The rapid and correct identification of pathogens is of paramount importance for the treatment of patients with invasive infections. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) can speed up the identification of bacteria and fungi and has quickly been embraced by medical microbiology laboratories worldwide. Different MALDI-TOF systems have been compared in studies focussing on identification rates of different pathogens. Another aspect that has not been systematically assessed is the performance in daily routine and handling, which is important especially for microbiology routine laboratories. We compared two widespread commercial systems, Microflex LT Biotyper (Bruker) and VitekMS (bioMérieux), for the identification of 210 relevant clinical yeasts under routine conditions, using a time-saving direct transfer protocol. We assessed the need for an additional extraction step, the threshold for species identification and the duration of measurements with the two systems. The tested yeasts included 34 Candida albicans isolates, 144 non-albicans Candida spp. and 32 yeasts of different genera. The results of the two MS systems were compared with that of biochemical identification and, in case of discrepancies, DNA sequencing of the internal transcribed spacer or the large subunit of ribosomal DNA. Both systems correctly identified 96.2 % of isolates [202/210, non-significant (n.s.)]. Misidentifications were observed for VitekMS only (n = 5, no major errors, n.s.). VitekMS was the slower system (19.8 vs. 8.0 min for 10 samples, p = 0.002) but had the advantage of a more effective direct transfer protocol with less need for an additional extraction step.

摘要

快速准确地鉴定病原体对于侵袭性感染患者的治疗至关重要。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)能够加速细菌和真菌的鉴定,已迅速被全球医学微生物实验室所采用。在针对不同病原体鉴定率的研究中,已对不同的MALDI-TOF系统进行了比较。另一个尚未得到系统评估的方面是日常工作中的性能和操作,这对于微生物学常规实验室尤为重要。我们使用一种省时的直接转移方案,比较了两种广泛使用的商业系统,即Microflex LT Biotyper(布鲁克公司)和VitekMS(生物梅里埃公司),在常规条件下对210株相关临床酵母的鉴定情况。我们评估了是否需要额外的提取步骤、物种鉴定的阈值以及这两种系统的测量时间。测试的酵母包括34株白色念珠菌分离株、144株非白色念珠菌属念珠菌和32株不同属的酵母。将这两种质谱系统的结果与生化鉴定结果进行比较,如有差异,则对核糖体DNA的内转录间隔区或大亚基进行DNA测序。两种系统均正确鉴定了96.2%的分离株[202/210,无显著性差异(n.s.)]。仅在VitekMS系统中观察到错误鉴定(n = 5,无重大错误,n.s.)。VitekMS系统速度较慢(10个样本分别为19.8分钟和8.0分钟,p = 0.002),但具有更有效的直接转移方案的优势,额外提取步骤的需求较少。

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