...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Clinical Microbiology >Rapid, Sensitive, and Specific Escherichia coli H Antigen Typing by Matrix-Assisted Laser Desorption Ionization–Time of Flight-Based Peptide Mass Fingerprinting
【24h】

Rapid, Sensitive, and Specific Escherichia coli H Antigen Typing by Matrix-Assisted Laser Desorption Ionization–Time of Flight-Based Peptide Mass Fingerprinting

机译：通过基质辅助激光解吸电离快速，灵敏和特异的大肠杆菌H抗原分型–飞行时间的肽质谱指纹图谱时间

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has gained popularity in recent years for rapid bacterial identification, mostly at the genus or species level. In this study, a rapid method to identify the Escherichia coli flagellar antigen (H antigen) at the subspecies level was developed using a MALDI-TOF MS platform with high specificity and sensitivity. Flagella were trapped on a filter membrane, and on-filter trypsin digestion was performed. The tryptic digests of each flagellin then were collected and analyzed by MALDI-TOF MS through peptide mass fingerprinting. Sixty-one reference strains containing all 53 H types and 85 clinical strains were tested and compared to serotyping designations. Whole-genome sequencing was used to resolve conflicting results between the two methods. It was found that DHB (2,5-dihydroxybenzoic acid) worked better than CHCA (α-cyano-4-hydroxycinnamic acid) as the matrix for MALDI-TOF MS, with higher confidence during protein identification. After method optimization, reference strains representing all 53 E. coli H types were identified correctly by MALDI-TOF MS. A custom E. coli flagellar/H antigen database was crucial for clearly identifying the E. coli H antigens. Of 85 clinical isolates tested by MALDI-TOF MS-H, 75 identified MS-H types (88.2%) matched results obtained from traditional serotyping. Among 10 isolates where the results of MALDI-TOF MS-H and serotyping did not agree, 60% of H types characterized by whole-genome sequencing agreed with those identified by MALDI-TOF MS-H, compared to only 20% by serotyping. This MALDI-TOF MS-H platform can be used for rapid and cost-effective E. coli H antigen identification, especially during E. coli outbreaks.
机译：近年来，基质辅助激光解吸电离飞行时间质谱仪（MALDI-TOF MS）已迅速普及，可用于细菌快速鉴定，主要是在属或种水平上。在这项研究中，使用具有高特异性和敏感性的MALDI-TOF MS平台，开发了一种在亚种水平上鉴定大肠杆菌鞭毛抗原（H抗原）的快速方法。将鞭毛捕集在滤膜上，并进行滤器内胰蛋白酶消化。然后收集每个鞭毛蛋白的胰蛋白酶消化物，并通过肽质量指纹图谱通过MALDI-TOF MS进行分析。测试了包含所有53个H型和85个临床菌株的61个参考菌株，并与血清分型名称进行了比较。全基因组测序用于解决两种方法之间的矛盾结果。已发现，DHB（2,5-二羟基苯甲酸）的效果优于CHCA（α-氰基-4-羟基肉桂酸）作为MALDI-TOF MS的基质，在蛋白质鉴定过程中具有更高的可信度。方法优化后，通过MALDI-TOF MS正确鉴定了代表所有53种大肠杆菌H型的参考菌株。定制的大肠杆菌鞭毛/ H抗原数据库对于清楚识别大肠杆菌H抗原至关重要。在通过MALDI-TOF MS-H测试的85种临床分离株中，有75种已鉴定的MS-H类型（88.2％）与传统血清分型获得的结果相符。在MALDI-TOF MS-H和血清分型结果不一致的10个分离株中，以全基因组测序为特征的H型与MALDI-TOF MS-H鉴定的分离株一致，而通过血清分型只有20％。此MALDI-TOF MS-H平台可用于快速且经济高效的大肠杆菌H抗原鉴定，尤其是在大肠杆菌暴发期间。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号