摘要
目的:探索建立一种新的出血性大肠杆菌O157∶H7(Escherichia coli O157∶H7)的快速定量检测技术。方法:以硝酸纤维素膜为基片,用生物芯片点样仪制作抗体宏阵列,采用'双抗夹心'原理,对出血性大肠杆菌O157∶H7进行定量检测研究。结果:本方法对出血性大肠杆菌O157∶H7的检出限为3.4×105 CFU/mL,在105~107CFU/mL细菌浓度范围内,宏阵列的灰度值与细菌浓度之间有较好的线性关系。该方法可在2.5 h内同步检测多个样品中出血性大肠杆菌O157∶H7的浓度。结论:通过用标准菌株、模拟带菌等研究显示,用宏阵列技术快速定量检测出血性大肠杆菌O157∶H7,结果肉眼可见,稳定准确,同时操作简便、成本低廉、无需大型设备,制作好的抗体宏阵列可用于快速评估食品中出血性大肠杆菌O157∶H7的污染状况,尤其适合于基层实验室进行快速高通量样品筛查。
Purpose: To establish a new rapid quantitative assay for detecting Escherichia coli O157:H7. Methods: Using nitrocellulose membrane as substrate, an antibody macroarray was fabricated with a biochip spotting robot, and then a double antibody sandwich method was developed for quantitative detection of E. coli O157:H7. Results: The limit of detection(LOD) for E. coli O157:H7 was 3.4 × 105 CFU/mL, and a good linear relationship was observed between grey value for the macroarray and bacterial concentration in the range of 105–107 CFU/mL. This method could allow simultaneous determination of different concentrations of E. coli O157:H7 in multiple samples. Conclusions: As evaluated by applying it to standard strains and simulated enrichment culture, the antibody macroarray-based method could provide stable and accurate results visible to naked eyes with simple operation and low costs without the use of large equipment. Moreover, the antibody macroarray developed in this study could be used to rapidly assess E. coli O157:H7 pollution in food, especially suitable for rapid screening of samples in grassroots-level laboratories.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2014年第20期185-191,共7页
Food Science
基金
国家质检总局科技项目(GSCIQ_2010IK220)
上海市科委重点支撑项目(13430502400)
甘肃省科技支撑计划项目(1304FKCA056)
