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Differentiation of Streptococcus pneumoniae Conjunctivitis Outbreak Isolates by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry▿

机译:基质辅助激光解吸电离-飞行时间质谱法鉴别肺炎链球菌结膜炎暴发分离株▿

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摘要

Streptococcus pneumoniae (pneumococcus [Pnc]) is a causative agent of many infectious diseases, including pneumonia, septicemia, otitis media, and conjunctivitis. There have been documented conjunctivitis outbreaks in which nontypeable (NT), nonencapsulated Pnc has been identified as the etiological agent. The use of mass spectrometry to comparatively and differentially analyze protein and peptide profiles of whole-cell microorganisms remains somewhat uncharted. In this report, we discuss a comparative proteomic analysis between NT S. pneumoniae conjunctivitis outbreak strains (cPnc) and other known typeable or NT pneumococcal and streptococcal isolates (including Pnc TIGR4 and R6, Streptococcus oralis, Streptococcus mitis, Streptococcus pseudopneumoniae, and Streptococcus pyogenes) and nonstreptococcal isolates (including Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus) as controls. cPnc cells and controls were grown to mid-log phase, harvested, and subsequently treated with a 10% trifluoroacetic acid-sinapinic acid matrix mixture. Protein and peptide fragments of the whole-cell bacterial isolate-matrix combinations ranging in size from 2 to 14 kDa were evaluated by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Additionally Random Forest analytical tools and dendrogramic representations (Genesis) suggested similarities and clustered the isolates into distinct clonal groups, respectively. Also, a peak list of protein and peptide masses was obtained and compared to a known Pnc protein mass library, in which a peptide common and unique to cPnc isolates was tentatively identified. Information gained from this study will lead to the identification and validation of proteins that are commonly and exclusively expressed in cPnc strains which could potentially be used as a biomarker in the rapid diagnosis of pneumococcal conjunctivitis.
机译:肺炎链球菌(肺炎球菌[Pnc])是许多传染病的病原,包括肺炎,败血病,中耳炎和结膜炎。已经有结膜炎暴发的报道,其中已将不可分型(NT),未包囊的Pnc鉴定为病原体。使用质谱分析法比较和差异分析全细胞微生物的蛋白质和肽谱仍然有些未知。在本报告中,我们讨论了NT肺炎链球菌结膜炎暴发株(cPnc)与其他已知的可分型或NT肺炎球菌和链球菌分离株(包括Pnc TIGR4和R6,口腔链球菌,口疮链球菌,链球菌和假性肺炎球菌, )和非链球菌分离株(包括大肠杆菌,粪肠球菌和金黄色葡萄球菌)作为对照。将cPnc细胞和对照培养至对数中期,收获,然后用10%三氟乙酸-芥子酸基质混合物处理。通过基质辅助激光解吸电离飞行时间质谱法评估了全细胞细菌分离物-基质组合的蛋白质和肽片段的大小,范围为2至14 kDa。此外,Random Forest分析工具和树状图表示法(Genesis)提出了相似性,并将分离物分别分为不同的克隆组。同样,获得了蛋白质和肽质量的峰列表,并将其与已知的Pnc蛋白质量文库进行比较,在该文库中初步确定了cPnc分离物共有和独特的肽。从这项研究中获得的信息将有助于鉴定和验证在cPnc菌株中通常唯一表达的蛋白质,这些蛋白质可能被用作快速诊断肺炎球菌性结膜炎的生物标记。

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