Baldauf Nathan A, Rodriguez-Romo Luis A, Männig Annegret, Yousef Ahmed E, Rodriguez-Saona Luis E
Department of Food Science and Technology, The Ohio State University 2015 Fyffe Road, Parker Food Science Building, Columbus, Ohio 43210, USA.
J Microbiol Methods. 2007 Jan;68(1):106-14. doi: 10.1016/j.mimet.2006.06.012. Epub 2006 Aug 14.
Salmonella enterica serovars are prevalent foodborne pathogens responsible for high numbers of salmonellosis each year. Complex Fourier-transform infrared (FTIR) spectra offer unique biochemical fingerprints of bacteria with bands due to major cellular components. Growth media effects on discrimination of Salmonella serovars by FTIR spectroscopy were investigated and a novel sample preparation technique was developed. S. enterica strains from six serovars were grown on xylose lysine desoxycholate (XLD), Miller-Mallinson (MM), and plate count (PCA) agar as a control (37 degrees C, 24 h). Isolated colonies were suspended in 50% acetonitrile and centrifuged; the remaining pellet was placed on an AMTIR (attenuated total reflectance) crystal and dried under vacuum. Classification models (Soft Independent Modeling of Class Analogy, SIMCA), generated from derivatized infrared spectra (1300-900 cm-1 or 1200-900 cm-1), successfully discriminated among Salmonella strains with major discrimination from 1000-970 cm-1 associated to stretching modes of O-specific polysaccharide chains of lipopolysaccharides. Sample treatment with acetonitrile enhanced safe handling of the bacteria, removed interfering signals and improved the discriminating ability of SIMCA. All media were able to discriminate the S. enterica strains studied, varying in discriminating peaks and class distances in SIMCA classification. This methodology, with the production of large libraries of pathogenic bacteria, could be applied for the rapid monitoring of bacterial contamination in food with minimal sample manipulation.
肠炎沙门氏菌血清型是常见的食源性病原体,每年导致大量沙门氏菌病。复杂的傅里叶变换红外(FTIR)光谱提供了细菌独特的生化指纹图谱,其谱带源于主要的细胞成分。研究了生长培养基对FTIR光谱鉴别沙门氏菌血清型的影响,并开发了一种新的样品制备技术。来自六个血清型的肠炎沙门氏菌菌株在木糖赖氨酸脱氧胆酸盐(XLD)、米勒 - 马林森(MM)和作为对照的平板计数(PCA)琼脂上培养(37摄氏度,24小时)。将分离的菌落悬浮于50%乙腈中并离心;将剩余的沉淀置于衰减全反射(AMTIR)晶体上并真空干燥。由衍生化红外光谱(1300 - 900 cm-1或1200 - 900 cm-1)生成的分类模型(类类比软独立建模,SIMCA)成功区分了沙门氏菌菌株,主要区分在1000 - 970 cm-1之间,这与脂多糖O - 特异性多糖链的拉伸模式有关。用乙腈进行样品处理增强了对细菌的安全处理,去除了干扰信号并提高了SIMCA的鉴别能力。所有培养基都能够区分所研究的肠炎沙门氏菌菌株,在SIMCA分类中的鉴别峰和类距离有所不同。这种方法,随着病原菌大型文库的建立,可用于在最少样品处理的情况下快速监测食品中的细菌污染。