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基质辅助激光解吸电离飞行时间质谱系统在快速鉴定具有公共卫生重要性的菌种中的应用。

Application of MALDI-TOF MS Systems in the Rapid Identification of spp. of Public Health Importance.

作者信息

Hsieh Ying-Hsin, Wang Yun F, Moura Hercules, Miranda Nancy, Simpson Steven, Gowrishankar Ramnath, Barr John, Kerdahi Khalil, Sulaiman Irshad M

机构信息

U.S. Food and Drug Administration, Microbiological Sciences Branch, Southeast Food and Feed Laboratory, 60 8th St NE, Atlanta, GA 30309.

Emory University School of Medicine, 1364 Clifton Rd NE, Atlanta, GA 30322; Grady Memorial Hospital, Clinical Pathology Laboratory, 80 Jesse Hill Dr, Atlanta, GA 30303.

出版信息

J AOAC Int. 2018 May 1;101(3):761-768. doi: 10.5740/jaoacint.17-0266. Epub 2017 Sep 12.

Abstract

Campylobacteriosis is an infectious gastrointestinal disease caused by Campylobacter spp. In most cases, it is either underdiagnosed or underreported due to poor diagnostics and limited databases. Several DNA-based molecular diagnostic techniques, including 16S ribosomal RNA (rRNA) sequence typing, have been widely used in the species identification of Campylobacter. Nevertheless, these assays are time-consuming and require a high quality of bacterial DNA. Matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) MS is an emerging diagnostic technology that can provide the rapid identification of microorganisms by using their intact cells without extraction or purification. In this study, we analyzed 24 American Type Culture Collection reference isolates of 16 Campylobacter spp. and five unknown clinical bacterial isolates for rapid identification utilizing two commercially available MADI-TOF MS platforms, namely the bioMérieux VITEK® MS and Bruker Biotyper systems. In addition, 16S rRNA sequencing was performed to confirm the species-level identification of the unknown clinical isolates. Both MALDI-TOF MS systems identified the isolates of C. jejuni, C. coli, C. lari, and C. fetus. The results of this study suggest that the MALDI-TOF MS technique can be used in the identification of Campylobacter spp. of public health importance.

摘要

弯曲杆菌病是一种由弯曲杆菌属引起的感染性胃肠道疾病。在大多数情况下,由于诊断手段不足和数据库有限,该病要么诊断不足,要么报告不足。包括16S核糖体RNA(rRNA)序列分型在内的几种基于DNA的分子诊断技术已广泛用于弯曲杆菌的菌种鉴定。然而这些检测方法耗时且需要高质量的细菌DNA。基质辅助激光解吸电离(MALDI)飞行时间(TOF)质谱是一种新兴的诊断技术,它可以通过使用完整细胞而无需提取或纯化来快速鉴定微生物。在本研究中,我们利用两个商用MADI-TOF MS平台,即生物梅里埃VITEK® MS和布鲁克Biotyper系统,分析了16种弯曲杆菌属的24株美国典型培养物保藏中心参考菌株和5株未知临床细菌分离株,以进行快速鉴定。此外,还进行了16S rRNA测序以确认未知临床分离株的菌种水平鉴定结果。两个MALDI-TOF MS系统均鉴定出空肠弯曲杆菌、大肠弯曲杆菌、海鸥弯曲杆菌和胎儿弯曲杆菌的分离株。本研究结果表明,MALDI-TOF MS技术可用于鉴定具有公共卫生重要性的弯曲杆菌属菌种。

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