欢迎来到艾兰博曼,请 登录 | 立即注册
查看: 886|回复: 2

【文献翻译】Mass Spectrometry–Based Escherichia coli H Antigen/Flagel...

[复制链接]

2

听众

1

收听

214

积分

版主

Rank: 7Rank: 7Rank: 7

积分
214
发表于 2016-6-2 15:07:22 | 显示全部楼层 |阅读模式
Mass Spectrometry–Based Escherichia coli H Antigen/Flagella Typing: Validation and Comparison with Traditional Serotyping

BACKGROUND: Escherichia coli H antigen typing with antisera, a useful method for flagella clinical identification and classification, is a time-consuming process because of the need to induce flagella growth and the occurrence of undetermined strains. We developed an alternative rapid and analytically sensitive mass spectrometry (MS) method, termed MS-based H antigen typing (MS-H), and applied it at the protein sequence level for H antigen typing. We also performed a comparison with traditional serotyping on reference strains and clinical isolates.

METHODS: On the basis of international guidelines, the analytical selectivity and sensitivity, imprecision, correlation, repeatability, and reproducibility of the MS-H platform was evaluated using reference strains. Comparison of MS-H typing and serotyping was performed using 302 clinical isolates from 5 Canadian provinces, and discrepant results between the 2 platforms were resolved through whole genome sequencing.

RESULTS: Repeated tests on reference strain EDL933 demonstrated a lower limit of the measuring interval at the subsingle colony (16.97 μg or 1.465 × 107 cells) level and close correlation (r2 > 0.99) between cell culture biomass and sequence coverage. The CV was <10.0% among multiple repeats with 4 reference strains. Intra- and interlaboratory tests demonstrated that the MS-H method was robust and reproducible under various sample preparation and instrumentation conditions. Using discrepancy analysis via whole genome sequencing, performed on isolates with discrepant results, MS-H accurately identified 12.3% more isolates than conventional serotyping.

CONCLUSIONS: MS-H typing of E. coli is useful for fast and accurate flagella typing and could be very useful during E. coli outbreaks.

链接:http://www.clinchem.org/content/62/6/839.abstract

0

听众

0

收听

48

积分

新手上路

Rank: 1

积分
48
发表于 2016-6-3 12:47:50 | 显示全部楼层
领了~~~~~~

0

听众

0

收听

48

积分

新手上路

Rank: 1

积分
48
发表于 2016-6-3 14:29:53 | 显示全部楼层
基于质谱的大肠杆菌H抗原/鞭毛分型:与传统血清分型的验证与比较

背景:通过抗血清进行的大肠杆菌H抗原分型是一种在鞭毛临床识别和分类中有用的方法,然而由于其需要诱导鞭毛生长和未确定的菌株的出现,是一种耗时的方法。我们开发了一种替代性的快速且分析灵敏的质谱(MS)方法,称为基于MS的H抗原分型(MS-H),并将其应用于H抗原分型的蛋白质序列级别。我们还就参考菌株和临床分离菌株进行了与传统的血清分型的比较。

方法:根据国际准则,使用参考菌株对MS-H平台分析的选择性和灵敏度、不精确性、相关性、重复性和的再现性进行评价。使用加拿大五个省的302株临床分离菌株对MS-H分型和血清分型进行比较,这两个平台之间不一致的结果通过全基因组测序解析。

结果:对参考菌菌株EDL933进行的重复试验,揭示了在亚单群落级别的测量间隔的下限较低(16.97微克或1.465×107个细胞)且细胞培养物生物量与序列覆盖率密切相关(R2> 0.99)。在针对四个参考菌株的多次重复实验中CV<10.0%。实验室内和实验室间测试表明,MS-H方法在不同的样品制备方式和仪器条件下的稳定且可重复。通过全基因组测序对存在不一致的结果的分离株进行差异分析,MS-H可以准确识别比传统的血清分型多12.3%的分离株。

结论:大肠杆菌的MS-H分型对于快速且准确进行鞭毛分型有用,并且在大肠杆菌爆发时非常有用。

链接:HTTP://www.clinchem.org/content/62/6/839.abstract
您需要登录后才可以回帖 登录 | 立即注册

本版积分规则

© 2013 艾兰博曼 All Rights Reserved.
( 浙ICP备07020270号-8 )
 
快速回复 返回顶部 返回列表