Raman spectroscopic typing reveals the presence of carotenolds in Mycoplasma pneumoniae
Raman spectroscopic typing reveals the presence of carotenolds in Mycoplasma pneumoniae
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DOI：
10.1099/mic.0.026724-0
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发表时间：
2009-06-01
期刊：
影响因子：
2.8
通讯作者：
Vink, Cornelis
中科院分区：
文献类型：
--
作者：
Maquelin, Kees;Hoogenboezem, Theo;Vink, Cornelis
拉曼光谱已被证明是一种对微生物进行鉴定和/或分型的非常有用的方法。在这项研究中，我们开始评估这项技术是否也可以用作区分肺炎支原体分离株的工具，肺炎支原体通常被认为是基因高度一致的物种。在这项评估中，共分析了104株肺炎支原体，包括两株参考菌株(M129和FH)和102株临床分离株，它们是1973年至2005年期间分离的，来自不同的国家。通过对该菌株的拉曼光谱分析(拉曼分型)，我们能够重复地区分六个不同的菌株群。拉曼分型和基于肺炎支原体P1(或MPN 141)基因序列差异的P1基因分型之间没有明显的相关性。在我们鉴定的两个主要拉曼簇(簇3和簇6，总共包含81%的菌株)中，不同的P1亚型分布相似，类似于76%的分离物属于亚型1，类似于亚型2的20%，类似于变异体2a的5%。然而，在第2组和第5组(1100%)以及第1组(75%)和第4组(71%)中发现了较高的P1亚型2毒株的流行率；然而，这些群组中含有少量的毒株。只有两个菌株(2%)不能正确分型。有趣的是，拉曼光谱分析显示肺炎支原体中存在类胡萝卜素。这一发现与肺炎支原体基因的鉴定是一致的，这些基因与导致类胡萝卜素合成的生化途径，即2-C-甲基-D-赤藓糖醇4-磷酸(MEP)途径有相似之处。因此，我们假设肺炎支原体存在一条类似MEP的类胡萝卜素合成途径。我们的结论是，拉曼光谱是一种方便的工具来区分肺炎支原体菌株，它是对目前这种细菌分型方法的一个有前途的补充。
Raman spectroscopy has previously been demonstrated to be a highly useful methodology for the identification and/or typing of micro-organisms. In this study, we set out to evaluate whether this technology could also be applied as a tool to discriminate between isolates of Mycoplasma pneumoniae, which is generally considered to be a genetically highly uniform species. In this evaluation, a total of 104 strains of M. pneumoniae were analysed, including two reference strains (strains M129 and FH), and 102 clinical isolates, which were isolated between 1973 and 2005 and originated from various countries. By Raman spectral analysis (Raman typing) of this strain collection, we were able to reproducibly distinguish six different clusters of strains. An unequivocal correlation between Raman typing and P1 genotyping, which is based on sequence differences in the P1 (or MPN 141) gene of M. pneumoniae, was not observed. In the two major Raman clusters that we identified (clusters 3 and 6, which together harboured 81 % of the strains), the different P1 subtypes were similarly distributed, and similar to 76% isolates were of subtype 1, similar to 20% of subtype 2 and similar to 5 % of variant 2a. Nevertheless, a relatively high prevalence of P1 subtype 2 strains was found in clusters 2 and 5 (1100 %), as well as in cluster 1 (75 %) and cluster 4 (71 %); these clusters, however, harboured a small number of strains. Only two of the strains (2 %) could not be typed correctly. Interestingly, analysis of the Raman spectra revealed the presence of carotenoids in M. pneumoniae. This finding is in line with the identification of M. pneumoniae genes that have similarity with genes involved in a biochemical pathway leading to carotenoid synthesis, i.e. the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. Therefore, we hypothesize that M. pneumoniae hosts an MEP-like pathway for carotenoid synthesis. We conclude that Raman spectroscopy is a convenient tool for discriminating between M. pneumoniae strains, and that it presents a promising supplement to the current methods for typing of this bacterium.