Fluorescence characterization of clinically-important bacteria.
Fluorescence characterization of clinically-important bacteria.
复制标题
DOI：
10.1371/journal.pone.0075270
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Muller JP
中科院分区：
文献类型：
--
作者：
Dartnell LR;Roberts TA;Moore G;Ward JM;Muller JP
医疗保健相关感染（HCAI/HAI）是住院期间对患者健康的重大威胁，并为后续治疗带来数十亿美元的额外费用。一种在HAI爆发发生之前在临床环境中检测细菌污染的有希望的方法是利用细胞分子的天然荧光用于手持式快速扫描监测仪器。先前的研究表明，基于荧光的检测对于食源性和环境微生物是敏感和有效的，甚至能够区分细胞类型，但这种强大的技术尚未在宏观尺度上部署，用于医疗机构污染的主要监测，以防止HAI。在这里，我们报告的实验数据的规格和设计这样一个基于荧光的检测仪器。我们的特点是完整的荧光响应的11个临床相关的细菌产生的激发-发射矩阵（EEMs）在宽波长范围内。此外，已经分析了病房中常见的并且可能导致病原体转移的许多表面和设备项目的背景荧光掩蔽来自污染物细菌的信号的潜在问题。其中包括床边扶手、护士呼叫按钮、血压袖带和病房电脑键盘，以及消毒清洁产品和超细纤维布。所有检查的细菌菌株均表现出与色氨酸相关的独特双峰荧光特征，未检测到其他细胞荧光团。因此，该荧光调查发现，来自280 nm处的激发源的340 nm的发射峰是用于检测细菌污染的靶的细胞荧光信号。分析的大多数材料提供了一个光谱窗口，通过它确实可以检测到细菌污染。发现了一些背景荧光掩盖细菌荧光的潜在问题，但在微纤维清洁布的情况下，成像技术可以在形态上区分杂散链和细菌污染。
Healthcare-associated infections (HCAI/HAI) represent a substantial threat to patient health during hospitalization and incur billions of dollars additional cost for subsequent treatment. One promising method for the detection of bacterial contamination in a clinical setting before an HAI outbreak occurs is to exploit native fluorescence of cellular molecules for a hand-held, rapid-sweep surveillance instrument. Previous studies have shown fluorescence-based detection to be sensitive and effective for food-borne and environmental microorganisms, and even to be able to distinguish between cell types, but this powerful technique has not yet been deployed on the macroscale for the primary surveillance of contamination in healthcare facilities to prevent HAI. Here we report experimental data for the specification and design of such a fluorescence-based detection instrument. We have characterized the complete fluorescence response of eleven clinically-relevant bacteria by generating excitation-emission matrices (EEMs) over broad wavelength ranges. Furthermore, a number of surfaces and items of equipment commonly present on a ward, and potentially responsible for pathogen transfer, have been analyzed for potential issues of background fluorescence masking the signal from contaminant bacteria. These include bedside handrails, nurse call button, blood pressure cuff and ward computer keyboard, as well as disinfectant cleaning products and microfiber cloth. All examined bacterial strains exhibited a distinctive double-peak fluorescence feature associated with tryptophan with no other cellular fluorophore detected. Thus, this fluorescence survey found that an emission peak of 340nm, from an excitation source at 280nm, was the cellular fluorescence signal to target for detection of bacterial contamination. The majority of materials analysed offer a spectral window through which bacterial contamination could indeed be detected. A few instances were found of potential problems of background fluorescence masking that of bacteria, but in the case of the microfiber cleaning cloth, imaging techniques could morphologically distinguish between stray strands and bacterial contamination.
登录
查看更多内容
影响因子：
12.8
作者：
Elliott, S.;Lead, J. R.;Baker, A.
通讯作者：
Baker, A.
影响因子：
12.6
作者：
Estes, C;Duncan, A;Powers, L
通讯作者：
Powers, L
影响因子：
2.7
作者：
Ammor, Mohammed Salim
通讯作者：
Ammor, Mohammed Salim
影响因子：
11.4
作者：
Cory, RM;McKnight, DM
通讯作者：
McKnight, DM
影响因子：
3
作者：
Alberts, JJ;Takács, M
通讯作者：
Takács, M