基于磁性纳米颗粒的化学发光酶免疫分析用于日本血吸虫抗体的检测

Chemiluminescent enzyme immunoassay based on magnetic nanoparticles for the detection of schistosoma japonicum antibody

该文提出了一种高灵敏的基于磁性纳米颗粒的化学发光酶免疫分析方法用于感染兔血清中日本血吸虫抗体的检测。日本血吸虫抗原通过N-(3-二甲基氨丙基)-N-乙基碳二亚胺盐(EDC)和N-羟基琥珀酰亚胺(NHS)固定在磁性纳米颗粒的表面,选择性地结合与富集血清中的日本血吸虫抗体,然后与辣根过氧化物酶标记的羊抗兔二抗夹心进行免疫分析。对免疫反应的实验条件如日本血吸虫抗原在磁性纳米颗粒表面的标记浓度,磁性纳米颗粒的用量,抗原与待测抗体的反应时间及辣根过氧化物酶标记二抗的浓度进行了优化。发光强度与感染兔血清的稀释比在1∶10000～1∶100的范围内成良好的线性关系,检测下限为1∶13442。

A novel chemiluminescent enzyme immunoassay based on magnetic nanoparticles （MNPs） has been developed for highly sensitive and rapid detection of sehistosoma japonicum antibody （SjAb） in infected rabbit serum （IRS）. Schistosoma japonieum antigen （SjAg） was immobilized onto MNPs using the N - （3 - dimethylaminopropy） - N^＋ - ethylcarbodiimidehydrochloride （EDC） and N - hydroxysueeinmide （NHS） . After reacting with SjAb, the horseradish peroxidase-conjugated goat anti -rabbit IgG（HRP- IgG） was bound to the surface of nanoparticles. Followed by the separation using a magnet and washing, luminol was then added to the antgen - antibody complex and the detection of luminescent intensity was carried out. Assay conditions were optimized, including the concentration of SjAg used for labeling, the amount of SjAg - MNPs, the reaction time of SjAg with SjAb, and the concentration of HRP - IgG. A good linear relationship was obtained between the luminescent intensity and the dilution ratio in the dilution range of 1：10 000 to 1：100 with a detection limit of 1：13 442 of dilution ratio.