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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。
In response to the needs of rapid and highly sensitive and specific detection of pathogens, a new method of pathogen detection was proposed based on the combination of magnetic immunosensing and adenosine triphosphate (ATP) bioluminescence. First of all, magnetic nanoparticles were used in combination with specific antibodies to prepare immunomagnetic nanoparticles to specifically capture the target. Then, the captured pathogen was cleaved and detected by ATP bioluminescence, and the concentration of the target substance was measured according to the light intensity. E. coli (O157 ... H7) was used as the detection target. The calibration curve was linear with a correlation coefficient of 0.943 and the total test time was 2h at a bacterial concentration of 10 ~ 108CFU / mL (CFU was colony forming unit). The sample was measured 12 times repeatedly, the precision reached 4.88%, with good repeatability.