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使用基质辅助激光解吸电离飞行时间质谱法从选择性增菌肉汤中检测单核细胞增生李斯特菌。

Detection of Listeria monocytogenes from selective enrichment broth using MALDI-TOF Mass Spectrometry.

作者信息

Jadhav Snehal, Sevior Danielle, Bhave Mrinal, Palombo Enzo A

机构信息

Environment and Biotechnology Centre, Faculty of Life and Social Sciences, Swinburne University of Technology, PO Box 218, Hawthorn 3122 Victoria, Australia.

bioMérieux Australia Pty Ltd, Unit 25 Parkview Business Centre, 1 Maitland Place, Baulkham Hills, NSW 2153, Australia.

出版信息

J Proteomics. 2014 Jan 31;97:100-6. doi: 10.1016/j.jprot.2013.09.014. Epub 2013 Sep 27.

Abstract

UNLABELLED

Conventional methods used for primary detection of Listeria monocytogenes from foods and subsequent confirmation of presumptive positive samples involve prolonged incubation and biochemical testing which generally require four to five days to obtain a result. In the current study, a simple and rapid proteomics-based MALDI-TOF MS approach was developed to detect L. monocytogenes directly from selective enrichment broths. Milk samples spiked with single species and multiple species cultures were incubated in a selective enrichment broth for 24h, followed by an additional 6h secondary enrichment. As few as 1 colony-forming unit (cfu) of L. monocytogenes per mL of initial selective broth culture could be detected within 30h. On applying the same approach to solid foods previously implicated in listeriosis, namely chicken pâté, cantaloupe and Camembert cheese, detection was achieved within the same time interval at inoculation levels of 10cfu/mL. Unlike the routine application of MALDI-TOF MS for identification of bacteria from solid media, this study proposes a cost-effective and time-saving detection scheme for direct identification of L. monocytogenes from broth cultures.This article is part of a Special Issue entitled: Trends in Microbial Proteomics.

BIOLOGICAL SIGNIFICANCE

Globally, foodborne diseases are major causes of illness and fatalities in humans. Hence, there is a continual need for reliable and rapid means for pathogen detection from food samples. Recent applications of MALDI-TOF MS for diagnostic microbiology focused on detection of microbes from clinical specimens. However, the current study has emphasized its use as a tool for detecting the major foodborne pathogen, Listeria monocytogenes, directly from selective enrichment broths. This proof-of-concept study proposes a detection scheme that is more rapid and simple compared to conventional methods of Listeria detection. Very low levels of the pathogen could be identified from different food samples post-enrichment in selective enrichment broths. Use of this scheme will facilitate rapid and cost-effective testing for this important foodborne pathogen.

摘要

未标记

用于从食品中初步检测单核细胞增生李斯特菌并随后确认疑似阳性样本的传统方法需要长时间培养和生化测试,通常需要四到五天才能获得结果。在本研究中,开发了一种基于蛋白质组学的简单快速的基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)方法,用于直接从选择性增菌肉汤中检测单核细胞增生李斯特菌。将接种了单一菌种和多种菌种培养物的牛奶样本在选择性增菌肉汤中培养24小时,然后再进行6小时的二次增菌。在30小时内即可检测到每毫升初始选择性肉汤培养物中低至1个单核细胞增生李斯特菌菌落形成单位(cfu)。将相同方法应用于先前与李斯特菌病有关的固体食品,即鸡肉酱、哈密瓜和卡门培尔奶酪,在接种水平为10cfu/mL时,可在相同时间间隔内实现检测。与将MALDI-TOF MS常规应用于从固体培养基中鉴定细菌不同,本研究提出了一种经济高效且省时的检测方案,用于直接从肉汤培养物中鉴定单核细胞增生李斯特菌。本文是名为《微生物蛋白质组学趋势》的特刊的一部分。

生物学意义

在全球范围内,食源性疾病是人类患病和死亡的主要原因。因此,持续需要可靠且快速的方法来从食品样本中检测病原体。MALDI-TOF MS最近在诊断微生物学中的应用主要集中于从临床标本中检测微生物。然而,本研究强调了其作为直接从选择性增菌肉汤中检测主要食源性病原体——单核细胞增生李斯特菌的工具的用途。这项概念验证研究提出了一种比传统李斯特菌检测方法更快、更简单的检测方案。在选择性增菌肉汤中富集后,可从不同食品样本中鉴定出极低水平的病原体。使用该方案将有助于对这种重要的食源性病原体进行快速且经济高效的检测。

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