Ravindranath Sandeep P, Mauer Lisa J, Deb-Roy Chitrita, Irudayaraj Joseph
Department of Agricultural and Biological Engineering and Bindley Bioscience Center, Purdue University, 225 South University Street, West Lafayette, Indiana 47907, USA.
Anal Chem. 2009 Apr 15;81(8):2840-6. doi: 10.1021/ac802158y.
Magnetic nanoparticles functionalized with anti-Escherichia coli O157:H7 or anti-Salmonella typhimurium antibodies that can specifically bind to their target organisms were used to isolate E. coli O157:H7 and S. typhimurium separately from a cocktail of bacteria and from food matrixes. The pathogens were then detected using label-free IR fingerprinting. The binding and detection protocol was first validated using a benchtop FT-IR spectrometer and then applied to a portable mid-IR spectrometer to enable this approach as a point-of-detection technology. Highly selective detection was achieved in less than 30 min at both species (E. coli O157:H7 vs S. typhimurium ) and strain (E. coli O157:H7 vs E. coli K12) levels in complex food matrixes (2% milk, spinach extract) with a detection limit of 10(4)-10(5) CFU/mL. The combined approach of functionalized magnetic nanoparticles and IR spectroscopy imparts specificity through spectroscopic fingerprinting and selectivity through species-specific antibodies with an in-built sample extraction step and could be applied in the field for on-site food-borne pathogen monitoring.
用抗大肠杆菌O157:H7或抗鼠伤寒沙门氏菌抗体功能化的磁性纳米颗粒,能够特异性结合其目标微生物,被用于从混合细菌和食品基质中分别分离大肠杆菌O157:H7和鼠伤寒沙门氏菌。然后使用无标记红外指纹识别技术检测病原体。该结合和检测方案首先使用台式傅里叶变换红外光谱仪进行验证,然后应用于便携式中红外光谱仪,以使这种方法成为一种检测点技术。在复杂食品基质(2%牛奶、菠菜提取物)中,在不到30分钟的时间内,在物种(大肠杆菌O157:H7与鼠伤寒沙门氏菌)和菌株(大肠杆菌O157:H7与大肠杆菌K12)水平上均实现了高选择性检测,检测限为10(4)-10(5) CFU/mL。功能化磁性纳米颗粒和红外光谱的联合方法通过光谱指纹识别赋予特异性,通过物种特异性抗体赋予选择性,并带有内置的样品提取步骤,可应用于现场食品传播病原体监测领域。