Chen Longyan, Mungroo Nawfal, Daikuara Luciana, Neethirajan Suresh
BioNano Laboratory, School of Engineering, University of Guelph, Guelph, ON, N1G 2W1 12, Canada.
J Nanobiotechnology. 2015 Jun 25;13:45. doi: 10.1186/s12951-015-0106-4.
Rapid detection and discrimination of bacteria for biomedical and food safety applications remain a considerable challenge. We report a label-free near infrared surface-enhanced Raman scattering (NIR-SERS) method for the discrimination of pathogenic bacteria from drinking water. The approach relies on the in situ synthesis of silver nanoparticles (Ag NPs) within the bacterial cell suspensions.
Pre-treatment of cells with Triton X-100 significantly improved the sensitivity of the assay. Using this method, we were able to discriminate several common pathogenic bacteria such as Escherichia coli, Pseudomonas aeruginosa, Methicillin-resistant Staphylococcus aureus (MRSA) and Listeria spp. A comparison of the SERS spectra allowed for the discrimination of two Listeria species, namely L. monocytogenes and L. innocua. We further report the application of the method to discriminate two MRSA strains from clinical isolates. The complete assay was completed in a span of 5 min.
The proposed analytical method proves to be a rapid tool for selective and label-free identification of pathogenic bacterium. Pre-treatment of bacterial cells with Triton X-100 resulted in new features on the SERS spectra, allowing for a successful discrimination of common disease related bacteria including E. coli, P. aeruginosa, Listeria and MRSA. We also demonstrate that the spectral features obtained using in situ synthesis of nanoparticles could be could be used to differentiate two species of listeria. By using L. innocua as a model sample, we found the limit of detection of our assay to be 10(3) CFU/mL. The method can selectively discriminate different bacterial species, and has a potential to be used in the development of point-of-care diagnostics with biomedical and food safety applications.
对于生物医学和食品安全应用而言,快速检测和鉴别细菌仍然是一项重大挑战。我们报告了一种用于鉴别饮用水中致病细菌的无标记近红外表面增强拉曼散射(NIR-SERS)方法。该方法依赖于在细菌细胞悬液中原位合成银纳米颗粒(Ag NPs)。
用Triton X-100对细胞进行预处理显著提高了检测的灵敏度。使用该方法,我们能够鉴别几种常见的致病细菌,如大肠杆菌、铜绿假单胞菌、耐甲氧西林金黄色葡萄球菌(MRSA)和李斯特菌属。通过比较SERS光谱,可以鉴别两种李斯特菌,即单核细胞增生李斯特菌和无害李斯特菌。我们还报告了该方法在鉴别临床分离株中的两种MRSA菌株的应用。整个检测在5分钟内完成。
所提出的分析方法被证明是一种用于选择性和无标记鉴定致病细菌的快速工具。用Triton X-100对细菌细胞进行预处理导致SERS光谱出现新特征,从而能够成功鉴别包括大肠杆菌、铜绿假单胞菌、李斯特菌和MRSA在内的常见疾病相关细菌。我们还证明,使用纳米颗粒原位合成获得的光谱特征可用于区分两种李斯特菌。以无害李斯特菌作为模型样品,我们发现该检测方法的检测限为10³CFU/mL。该方法可以选择性地鉴别不同的细菌种类,并有可能用于开发具有生物医学和食品安全应用的即时诊断。