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鸡肉、梁和弯曲杆菌:通过振动光谱和基质辅助激光解吸电离质谱法对食源细菌进行快速鉴别

Chicken, beams, and Campylobacter: rapid differentiation of foodborne bacteria via vibrational spectroscopy and MALDI-mass spectrometry.

作者信息

Muhamadali Howbeer, Weaver Danielle, Subaihi Abdu, AlMasoud Najla, Trivedi Drupad K, Ellis David I, Linton Dennis, Goodacre Royston

机构信息

School of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.

出版信息

Analyst. 2016 Jan 7;141(1):111-22. doi: 10.1039/c5an01945a. Epub 2015 Nov 2.

Abstract

Campylobacter species are one of the main causes of food poisoning worldwide. Despite the availability of established culturing and molecular techniques, due to the fastidious nature of these microorganisms, simultaneous detection and species differentiation still remains challenging. This study focused on the differentiation of eleven Campylobacter strains from six species, using Fourier transform infrared (FT-IR) and Raman spectroscopies, together with matrix-assisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS), as physicochemical approaches for generating biochemical fingerprints. Cluster analysis of data from each of the three analytical approaches provided clear differentiation of each Campylobacter species, which was generally in agreement with a phylogenetic tree based on 16S rRNA gene sequences. Notably, although C. fetus subspecies fetus and venerealis are phylogenetically very closely related, using FT-IR and MALDI-TOF-MS data these subspecies were readily differentiated based on differences in the lipid (2920 and 2851 cm(-1)) and fingerprint regions (1500-500 cm(-1)) of the FT-IR spectra, and the 500-2000 m/z region of the MALDI-TOF-MS data. A finding that was further investigated with targeted lipidomics using liquid chromatography-mass spectrometry (LC-MS). Our results demonstrate that such metabolomics approaches combined with molecular biology techniques may provide critical information and knowledge related to the risk factors, virulence, and understanding of the distribution and transmission routes associated with different strains of foodborne Campylobacter spp.

摘要

弯曲杆菌属是全球食物中毒的主要原因之一。尽管已有成熟的培养和分子技术,但由于这些微生物的苛求特性,同时检测和菌种鉴别仍然具有挑战性。本研究聚焦于使用傅里叶变换红外光谱(FT-IR)、拉曼光谱以及基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)作为生成生化指纹图谱的物理化学方法,对来自六个菌种的11株弯曲杆菌菌株进行鉴别。对这三种分析方法各自的数据进行聚类分析,能够清晰地区分每种弯曲杆菌菌种,这与基于16S rRNA基因序列构建的系统发育树总体一致。值得注意的是,尽管胎儿弯曲杆菌胎儿亚种和性病亚种在系统发育上密切相关,但利用FT-IR和MALDI-TOF-MS数据,基于FT-IR光谱中脂质区域(2920和2851 cm(-1))和指纹区域(1500 - 500 cm(-1))的差异以及MALDI-TOF-MS数据的500 - 2000 m/z区域,可轻松区分这两个亚种。这一发现通过使用液相色谱 - 质谱联用(LC-MS)的靶向脂质组学进一步研究。我们的结果表明,这种代谢组学方法与分子生物学技术相结合,可能提供与食源性病原体弯曲杆菌不同菌株相关的风险因素、毒力以及对其分布和传播途径理解的关键信息和知识。

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