Baldauf Nathan A, Rodriguez-Romo Luis A, Yousef Ahmed E, Rodriguez-Saona Luis E
Department of Food Science and Technology, The Ohio State University, Parker Food Science Building, 2015 Fyffe Road, Columbus, OH 43210, USA.
Appl Spectrosc. 2006 Jun;60(6):592-8. doi: 10.1366/000370206777670521.
Salmonella enterica serovars include pathogens responsible for high numbers of foodborne salmonellosis. Fourier transform infrared (FT-IR) spectroscopy can be used to rapidly and accurately identify microorganisms based on unique spectra of bacterial cell components. The objectives of this study were to discriminate closely related Salmonella enterica serovars by using FT-IR spectroscopy and multivariate analysis and to compare the performance of three techniques for differentiating among Salmonella serovars. Selected serovars of S. enterica were streaked onto plate count agar and incubated (37 degrees C, 24 h). Isolated colonies were suspended in phosphate buffer or 50% ethanol (10 microL). Suspensions were placed on (1) ZnSe crystals for transmission, (2) disposable polyethylene membranes (DPM) for transmission, and (3) diamond crystal plate for attenuated total reflectance (ATR) analyses; all samples were dried under vacuum. Classification models, soft independent modeling of class analogy (SIMCA), from derivatized infrared spectra (1300-900 cm(-1)), discriminated among Salmonella serovars presumably attributed to cell's lipopolysaccharides (1000-980 cm(-1)). Samples on DPM required high cell density for reliable spectra. High-quality spectra were obtained when a single colony was suspended in ethanol or buffer and mounted on ZnSe crystals for transmission or diamond plate for ATR analysis. Prediction of unknowns, representative of serovars used to construct classification models, showed that all techniques were suitable for the rapid and accurate differentiation of Salmonella serovars.
肠炎沙门氏菌血清型包括导致大量食源性沙门氏菌病的病原体。傅里叶变换红外(FT-IR)光谱可用于基于细菌细胞成分的独特光谱快速准确地鉴定微生物。本研究的目的是利用FT-IR光谱和多变量分析来区分密切相关的肠炎沙门氏菌血清型,并比较三种区分沙门氏菌血清型技术的性能。将选定的肠炎沙门氏菌血清型划线接种到平板计数琼脂上并培养(37℃,24小时)。将分离的菌落悬浮于磷酸盐缓冲液或50%乙醇(10微升)中。悬浮液分别置于(1)用于透射的ZnSe晶体上、(2)用于透射的一次性聚乙烯膜(DPM)上以及(3)用于衰减全反射(ATR)分析的金刚石晶体板上;所有样品均在真空下干燥。从衍生化红外光谱(1300 - 900厘米-1)建立的分类模型,即类类比软独立建模(SIMCA),可区分沙门氏菌血清型,推测这归因于细胞的脂多糖(1000 - 980厘米-1)。置于DPM上的样品需要高细胞密度才能获得可靠的光谱。当将单个菌落悬浮于乙醇或缓冲液中并置于用于透射的ZnSe晶体上或用于ATR分析的金刚石板上时,可获得高质量的光谱。对用于构建分类模型的血清型未知样品的预测表明,所有技术都适用于快速准确地区分沙门氏菌血清型。