基于磁免疫/生物发光的病原体快速检测研究

Pathogeny Rapid Detection Based on Immunomagnetic Separation and Biolumescence

针对病原体快速且高灵敏度和特异性检测的需求,将磁免疫技术与三磷酸腺苷(ATP)生物发光原理结合,提出一种新的致病菌检测方法。首先,采用磁性纳米粒子与特异性抗体结合制备免疫磁性纳米探针,特异性捕获目标物。然后,采用ATP生物发光技术对捕获的病原体裂解并检测,根据光强测量目标物浓度。以大肠杆菌(O157…H7)作为检测对象,在细菌浓度为10～108CFU/mL(CFU为菌落形成单位)范围内,该方法与传统培养法线性相关系数为0.943,总测试时间为2h。对样品重复测量12次,精密度达到4.88%,具有较好的重复性。

Focus on the requirment of pathogeny detection with high speed,high sensitivity and specificity,a novel method integrated with immunomagnetic separation and adenosine triphosphate（ATP） bioluminescence was developed.Firstly,an immunomagnetic nano probe was prepared by conjugating nano magnetic particles with antibody to capture the aim pathogeny.Sencondly,the pathogeny captured by the nano probe was lysed and the ATP was detected by biolumescence.Escherichia coli（O157：H7） samples with concentration of 10-10^8 CFU/mL（CFU：colony-forming units） were detected.The results show a linear response to the traditional plate count method with a correlation coefficient of 0.934 within 2 h.The reproducibility was also tested by 12 samples with the same concentration of 103 CFU/mL and the coefficient of variabilty is 4.88%.