[MALDI质谱法诊断细菌种群相变的适用性]。

Mikrobiologiia Pub Date : 2015-05-01
N A Kryazhevskikh, N G Loiko, E V Demkina, A L Mulyukin, A T Lebedev, A M Gaponov, A V Tutel'yan, Yu A Nikolaeva, G I El'-Registan
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引用次数: 0

摘要

研究了MALDI质谱法区分表型期变异(菌落形态和毒力/无毒)的效率。5属腐养致病菌和机会致病菌(不动杆菌、节杆菌、红球菌、棒状杆菌和埃希氏菌)。MALDI光谱(在SA和HCCA基质上)的分析包括:(1)确定蛋白质光谱的相似性(共同蛋白峰占蛋白质总量的百分比),这反映了对象的系统发育关系,并被推荐用于近缘物种的鉴定;(2)共同峰强度比较;(3)作为变异的决定性特征的特定峰的存在。在标准分析条件下,MALDI图谱的相似性在属-种-株-变异行中呈增加趋势。共同峰的强度评估最适用于相变体之间的区分,特别是在其剖面高度相似的情况下。在不同培养基(LB琼脂、TSA和TGYg)上生长的具有相似菌落形态(S、R、M和S(M))的相变菌株(A. oxydans菌株K14)的蛋白质谱存在差异,反映了其生理特性的差异。这一发现与我们先前筛选的菌落形态相似,但对氯代苯酚分解的底物特异性不同的不透明棘菌的结果一致。MALDI光谱分析可能是检测这种变异的唯一有效方法。
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[Applicability of MALDI Mass Spectrometry for Diagnostics of Phase Variants in Bacterial Populations].

Efficiency of MALDI mass spectrometry for differentiation between phenotypic phase variants (in colony morphology and virulence/avirulence) was investigated.for saprotrophic and opportunistically pathogenic bacteria of five genera (Acinetobacter, Arthrobacter, Rhodococcus, Corynebacterium, and Escherichia). Analysis of MALDI spectra (on the SA and HCCA matrices) included: (1) determination of similarity of the protein spectra as a percentage of the common protein peaks to the total amount of proteins, which reflects the phylogenetic relationships of the objects and has been recommended for identification of closely related species; (2) comparison of intensities of the common peaks; and (3) the presence of specific peaks as determinative characteristics of the variants. Under the standard analytical conditions the similarity between the MALDI profiles was shown to increase in the row: genus-species-strain-variant. Assessment of intensities of the common peaks was most applicable for differentiation between phase variants, especially in the case of high similarity of their profiles. Phase variants (A. oxydans strain K14) with similar colony morphotypes (S, R, M, and S(m)) grown on different media (LB agar, TSA, and TGYg) exhibited differences in their protein profiles reflecting the differences in their physiological characteristics. This finding is in agreement with our previous results on screening of the R. opacus with similar colony morphology and different substrate specificity in decomposition of chlorinated phenols. Analysis of MALDI spectra is probably the only efficient method for detection of such variants.

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