大肠杆菌特异性快速检测系统研制

结合三磷酸腺苷(ATP)生物发光原理和免疫磁分离技术,研制了一种细菌特异性快速检测系统,可实现对细菌的特异性高灵敏度快速检测.该仪器通过改进光学检测模块的避光设计和放大采集电路的噪声抑制设计,及严格控制试剂反应条件,降低了检测背景,从而达到了更低的检测下限并提高了检测灵敏度.对大肠肝菌(ATCC25922)标准菌溶液进行检测,检测下限达到18 CFU/mL, 与培养计数法检测结果相关系数R达到0.993,选取一份样品进行重复性检测,相对平均偏差(R.A.D.)为6.48 %,变异系数(CV)为9.33 %.结合免疫磁分离技术,对混入的高浓度金黄色葡萄球菌的大肠杆菌溶液进行检测,检测结果与原始大肠肝菌培养计数结果一致,证明该仪器及方法对检测大肠肝菌具有良好的特异性.该系统检测速度快,准确度较高,重复性良好,特异性好,可用于食品及环境中的致病菌快速检测.

Development of an Instrument for Specific and Rapid Detection of Escherichia coli

Combination of adenosine triphosphate (ATP) bioluminescence principles and immune magnetic separation technology, an instrument for specific rapid detection of bacteria is presented in this paper, which can achieve high specificity of bacteria rapid detection sensitivity. By improving light shielding design of optical detecting module and noise suppression design of amplification and A/D circuit, strict controlling of reaction conditions, the detection background was reduced, so as to achieve a lower detection limit and improve the detection sensitivity. E.coli (ATCC25922) standard solutions were tested in the experiment by the system, and the detection limit reached 18CFU/mL.Compared with the result obtained by the standard aerobic plate count APC method, the correlation coefficient (r) is 0.993. And the the reproducibility of the system was tested. The relative average deviation (R.A.D.) is 6.48% and the coefficient of variability (CV) is 9.33%. Combined with immunomagnetic separation techniques, E. coli solutions mixed with high concentrations of Staphylococcus aureus (ATCC25923) were tested. The linear correlation coefficient between the results detected by the system and the APC method is 0.9571, indicating that this apparatus and method has a good specificity. The system is rapid, specific, accurate and repetitive, which can be used in rapid detection of pathogens in food and environment.