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Identification of Photorhabdus symbionts by MALDI-TOF MS

机译:通过MALDI-TOF质谱鉴定Photorhabdus共生体

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摘要

Species of the bacterial genus Photorhabus live in a symbiotic relationship with Heterorhabditis entomopathogenic nematodes. Besides their use as biological control agents against agricultural pests, some Photorhabdus species are also a source of natural products and are of medical interest due to their ability to cause tissue infections and subcutaneous lesions in humans. Given the diversity of Photorhabdus species, rapid and reliable methods to resolve this genus to the species level are needed. In this study, we evaluated the potential of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of Photorhabdus species. To this end, we established a collection of 54 isolates consisting of type strains and multiple field strains that belong to each of the validly described species and subspecies of this genus. Reference spectra for the strains were generated and used to complement a currently available database. The extended reference database was then used for identification based on the direct transfer sample preparation method and the protein fingerprint of single colonies. High-level discrimination of distantly related species was observed. However, lower discrimination was observed with some of the most closely related species and subspecies. Our results therefore suggest that MALDI-TOF MS can be used to correctly identify Photorhabdus strains at the genus and species level, but has limited resolution power for closely related species and subspecies. Our study demonstrates the suitability and limitations of MALDI-TOF-based identification methods for assessment of the taxonomic position and identification of Photorhabdus isolates.
机译:细菌属 Photorhabus 的物种与异横纹虫致病性线虫共生关系。除了用作针对农业害虫的生物防治剂外,一些光斑物种也是天然产物的来源,并且由于它们能够引起人类组织感染和皮下病变而具有医学意义。鉴于Photorhabdus物种的多样性,需要快速可靠的方法来将该属解析到物种水平。在这项研究中,我们评估了基质辅助激光解吸/电离飞行时间质谱法 (MALDI-TOF MS) 在鉴定光弹物种方面的潜力。为此,我们建立了一个由类型菌株和多个田间菌株组成的 54 个分离株的集合,这些菌株属于该属的每个有效描述的物种和亚种。生成菌株的参考谱图,并用于补充当前可用的数据库。然后利用扩展参考数据库,基于直接转移样品制备方法和单菌落的蛋白质指纹图谱进行鉴定。观察到对远缘物种的高度歧视。然而,对一些最密切相关的物种和亚种的歧视程度较低。因此,我们的结果表明,MALDI-TOF MS可用于在属和种水平上正确识别Photorhabdus菌株,但对密切相关的物种和亚种的分辨率有限。我们的研究证明了基于MALDI-TOF的鉴定方法在评估光弹分离株的分类位置和鉴定方面的适用性和局限性。

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