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【文献翻译】A Novel Peptidomic Approach to Strain Typing of Clinical Ac...

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发表于 2016-6-2 15:24:16 | 显示全部楼层 |阅读模式
A Novel Peptidomic Approach to Strain Typing of Clinical Acinetobacter baumannii Isolates Using Mass Spectrometry

BACKGROUND: Acinetobacter baumannii is a common nosocomial pathogen and strain-typing methods play an important role in hospital outbreak investigations and epidemiologic surveillance. We describe a method for identifying strain-specific peptide markers based on LC-MS/MS profiling of digested peptides. This method classified a test set of A. baumannii isolates collected from a hospital outbreak with discriminatory performance exceeding that of MALDI-TOF mass spectrometry.

METHODS: Following the construction of a species “pan-peptidome” by in silico translation and digestion of whole genome sequences, a hypothetical set of genome-specific peptides for an isolate was constructed from the disjoint set of the pan-peptidome and the isolate's calculated peptidome. The genome-specific peptidome guided selection of highly expressed genome-specific peptides from LC-MS/MS experimental profiles as potential peptide markers. The species specificity of each experimentally identified genome-specific peptide was confirmed through a Unipept lowest common ancestor analysis.

RESULTS: Fifteen A. baumannii isolates were analyzed to derive a set of genome- and species-specific peptides that could be used as peptide markers. Identified peptides were cross-checked with protein BLAST against a set of 22 A. baumannii whole genome sequences. A subset of these peptide markers was confirmed to be present in the actual peptide profiles generated by multiple reaction monitoring and targeted LC-MS/MS. The experimentally identified peptides separated these isolates into 6 strains that agreed with multilocus sequence typing analysis performed on the same isolates.

CONCLUSIONS: This approach may be generalizable to other bacterial species, and the peptides may be useful for rapid MS strain tracking of isolates with broad application to infectious disease diagnosis.

链接:http://www.clinchem.org/content/62/6/866.abstract

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发表于 2016-6-2 15:51:50 | 显示全部楼层
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发表于 2016-6-2 23:53:13 | 显示全部楼层
利用质谱临床分离鲍曼不动杆菌通过菌株分型上新的多肽表达方式
背景:鲍氏不动杆菌是一种常见的医院内病原菌,菌株分型方法在住院疫情调查和流行病学监测中发挥重要作用。作者描述的测量方法,利用LC-MS/MS(液-质连用/质谱上)确定的消化肽谱图鉴定菌株特异性肽标志物。该方法分类测试鉴别来自医院疫情的鲍氏不动杆菌分离株的性能优于MALDI-TOF (基质辅助激光解析串联飞行时间) 质谱。

方法:通过全基因组序列的分解和模拟翻译的建设一种“泛肽”, 基因组特异肽的一组假说表面隔离是由不相交集的泛肽和多肽组一组分离组成。基因组的特定应用导向以LC-MS/MS实验简介高表达基因组特异性肽并作为潜在的肽标记。每个实验确定的基因组特异性肽的特异性通过Unipept最低常用的原始分析。

结果:对十五株鲍氏不动杆菌进行分析,研究者得出一套基因组和物种特异性肽,该肽可作为多肽标记。利用蛋白BLAST交叉确定肽与一组22个鲍氏不动杆菌全基因组序列的不同。这些肽标记的子集被证实在多反应监测和有针对性的LC-MS/MS真实产生的肽谱。实验鉴定的多肽的分离菌株为6株,与多位点序列分型分析在同一分离株上。

结论:该方法可推广到其他种类的细菌,同时肽可用于快速质谱跟菌株分离并广泛应用于传染病的诊断。
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