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目的:建立定量检测人血清IL-37的双抗夹心ELISA。方法:以鼠抗人IL-37单抗作为捕获抗体,制备的兔抗人IL-37多抗作为检测抗体,HRP标记山羊抗兔Ig G为二抗,重组人IL-37蛋白为标准品,建立检测人IL-37的双抗夹心ELISA方法。并对该方法的工作条件进行优化,对其灵敏度、线性范围、重复性和对登革热非结构蛋白NS1阳性患者血清IL-37的检测效果进行评价。结果:重组IL-37蛋白为标准品建立的双抗夹心ELISA法检测灵敏度为1.465μg/L,线性范围为1.465~46.875μg/L,批内和批间变异系数分别为6.6%和11.7%。采用此方法对诊断为登革热的患者血清进行检测,结果显示非结构蛋白NS1阳性患者IL-37水平显著高于健康人对照组。结论:成功建立了双抗夹心ELISA检测方法,可用于人血清中IL-37的检测。
Objective: To establish a double-antibody sandwich ELISA for the quantitative determination of human serum IL-37. Methods: The anti-human IL-37 monoclonal antibody was used as the capture antibody. The rabbit anti-human IL-37 polyclonal antibody was used as the detection antibody. HRP-labeled goat anti-rabbit IgG was used as the secondary antibody and recombinant human IL-37 protein as the standard. To establish a double-antibody sandwich ELISA method for detecting human IL-37. The working conditions of this method were optimized and its sensitivity, linearity, repeatability and detection of serum IL-37 in non-structural dengue NS1-positive patients were evaluated. Results: The sensitivity of double-antibody sandwich ELISA assay was 1.465μg / L and the linear range was 1.465 ~ 46.875μg / L. The coefficient of variation (CV) between intra-assay and intra-assay was 6.6% and 11.7% respectively. Using this method to detect the serum of patients diagnosed with dengue, the results showed that the non-structural protein NS1-positive patients IL-37 levels were significantly higher than the healthy control group. Conclusion: The double-antibody sandwich ELISA assay has been successfully established and can be used to detect IL-37 in human serum.