Rodriguez-Saona L E, Khambaty F M, Fry F S, Dubois J, Calvey E M
Joint Institute for Food Safety and Applied Nutrition--University of Maryland, College Park, Maryland 20742, USA.
J Food Prot. 2004 Nov;67(11):2555-9. doi: 10.4315/0362-028x-67.11.2555.
The use of Fourier transform-near infrared (FT-NIR) spectroscopy combined with multivariate pattern recognition techniques was evaluated to address the need for a fast and senisitive method for the detection of bacterial contamination in liquids. The complex cellular composition of bacteria produces FT-NIR vibrational transitions (overtone and combination bands), forming the basis for identification and subtyping. A database including strains of Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Bacillus cereus, and Bacillus thuringiensis was built, with special care taken to optimize sample preparation. The bacterial cells were treated with 70% (vol/vol) ethanolto enhance safe handling of pathogenic strains and then concentrated on an aluminum oxide membrane to obtain a thin bacterial film. This simple membrane filtration procedure generated reproducible FT-NIR spectra that allowed for the rapid discrimination among closely related strains. Principal component analysis and soft independent modeling of class analogy of transformed spectra in the region 5,100 to 4,400 cm(-1) were able to discriminate between bacterial species. Spectroscopic analysis of apple juices inoculated with different strains of E. coli at approximately 10(5) CFU/ml showed that FT-NIR spectralfeatures are consistent with bacterial contamination and soft independent modeling of class analogy correctly predicted the identity of the contaminant as strains of E. coli. FT-NIR in conjunction with multivariate techniques can be used for the rapid and accurate evaluation of potential bacterial contamination in liquids with minimal sample manipulation, and hence limited exposure of the laboratory worker to the agents.
评估了傅里叶变换近红外(FT-NIR)光谱法与多元模式识别技术的结合使用,以满足对液体中细菌污染进行快速、灵敏检测方法的需求。细菌复杂的细胞组成会产生FT-NIR振动跃迁(泛音和组合带),构成识别和亚型分类的基础。建立了一个包含大肠杆菌、铜绿假单胞菌、枯草芽孢杆菌、蜡样芽孢杆菌和苏云金芽孢杆菌菌株的数据库,并特别注意优化样品制备。细菌细胞用70%(体积/体积)乙醇处理,以加强对致病菌株的安全处理,然后浓缩在氧化铝膜上以获得薄细菌膜。这种简单的膜过滤程序产生了可重复的FT-NIR光谱,能够快速区分密切相关的菌株。对5100至4400 cm(-1)区域变换光谱进行主成分分析和类类比软独立建模,能够区分细菌种类。对接种了约10(5) CFU/ml不同大肠杆菌菌株的苹果汁进行光谱分析表明,FT-NIR光谱特征与细菌污染一致,类类比软独立建模正确预测了污染物为大肠杆菌菌株。FT-NIR结合多元技术可用于对液体中潜在细菌污染进行快速、准确的评估,样品处理最少,从而使实验室工作人员接触这些病原体的机会有限。