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验证 Colibrí 仪器在自动化制备 MALDI-TOF MS 靶标用于酵母鉴定中的应用。

Validation of the Colibrí Instrument for Automated Preparation of MALDI-TOF MS Targets for Yeast Identification.

机构信息

Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Department of Microbiology and Infection Control, Brussels, Belgium.

出版信息

J Clin Microbiol. 2022 Jul 20;60(7):e0023722. doi: 10.1128/jcm.00237-22. Epub 2022 Jun 15.

DOI:10.1128/jcm.00237-22
PMID:35703578
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9297811/
Abstract

Recently, Copan (Italy) introduced the Colibrí instrument for automated colony picking and preparation of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) target plates. Our study aimed to validate this system for yeasts as such testing has not been performed yet and is a missing link needed to implement the system for routine use. Fifty-five strains were selected to evaluate the accuracy of Colibrí. For each strain, a sheep blood agar plate supplemented with X and V factors (HEM) and a Sabouraud agar plate (SAB) were inoculated and incubated using the WASPlab specimen processing system (Copan). After 18 h and 36 h of incubation, the isolates were spotted in parallel using Colibrí and manually onto MALDI-TOF target plates with the addition of formic acid and identified using MALDI-TOF mass spectrometry. The reproducibility was evaluated using ATCC reference and clinical isolate-derived strains. The cumulative percentage of acceptable identification scores (IDs) after 36 h was 91% for strains cultured on HEM plates using both Colibrí and the manual method. The SAB plates showed inferior results for both Colibrí (76%) and the manual method (78%). We observed an overall agreement of 92% at 18 h for identification of the strains on the HEM plates between Colibrí and the manual method and 94% after 36 h. For the SAB plates, the agreement was 78% after 18 h and 84% after 36 h. Apart from Candida dubliniensis and Candida tropicalis, all species were identified with 100% accuracy using Colibrí on HEM plates. We observed good agreement between Colibrí and the manual reference method. These results demonstrate that Colibrí is a reliable system for MALDI-TOF target preparation for yeast identification, allowing increased standardization and less hands-on time.

摘要

最近,意大利的 Copan 公司推出了用于自动化集落挑取和基质辅助激光解吸电离飞行时间(MALDI-TOF)靶板制备的 Colibrí 仪器。我们的研究旨在验证该系统对酵母的适用性,因为之前尚未对其进行测试,这是实施该系统进行常规使用所必需的缺失环节。我们选择了 55 株 菌株来评估 Colibrí 的准确性。对于每种菌株,在绵羊血琼脂板(添加 X 和 V 因子的 HEM)和沙氏琼脂板(SAB)上接种并使用 WASPlab 标本处理系统(Copan)孵育。孵育 18 小时和 36 小时后,使用 Colibrí 和手动方法将分离株同时点样到 MALDI-TOF 靶板上,加入甲酸,并使用 MALDI-TOF 质谱进行鉴定。使用 ATCC 参考菌株和临床分离株衍生的菌株评估了重复性。孵育 36 小时后,使用 Colibrí 和手动方法在 HEM 平板上培养的菌株的可接受鉴定评分(ID)的累积百分比为 91%。对于 Colibrí(76%)和手动方法(78%),SAB 平板的结果较差。在 18 小时时,HEM 平板上的菌株鉴定中,Colibrí 和手动方法之间的总体一致性为 92%,36 小时后为 94%。对于 SAB 平板,18 小时时的一致性为 78%,36 小时后为 84%。除了杜氏念珠菌和热带念珠菌外,使用 Colibrí 在 HEM 平板上对所有 种都进行了 100%准确的鉴定。我们观察到 Colibrí 与手动参考方法之间具有良好的一致性。这些结果表明,Colibrí 是一种用于酵母鉴定的 MALDI-TOF 靶板制备的可靠系统,可提高标准化程度并减少手动操作时间。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d20/9297811/788f6d626e26/jcm.00237-22-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d20/9297811/d9fcb87c32b6/jcm.00237-22-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d20/9297811/788f6d626e26/jcm.00237-22-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d20/9297811/d9fcb87c32b6/jcm.00237-22-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d20/9297811/788f6d626e26/jcm.00237-22-f002.jpg

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