基于SPR免疫传感器技术的登革热血清学检测方法研究

Serological detection method based on surface plasmon resonance immunosensor for dengue fever

目的建立一种非标记、多指标的登革热快速检测方法,应用于登革热疑似病例的血清样本的快速检测。方法采用表面引发聚合反应技术构建基于寡聚乙二醇高分子的抗蛋白质非特异性吸附表面,以抗登革热IgM、IgG和抗登革热NS1抗体分别作为捕获探针,建立登革热的多指标微阵列芯片。采用表面等离子共振检测技术的技术手段,在血清中检测登革热特异性抗体。结果该技术方法对262份登革热疑似病例血清进行检测,阳性检出率为6.9%(18/262),高于PCR 2.3%(6/262)和ELISA 5.3%(14/262),其中PCR阳性的6份血清样本均检测到NS1标记物,而14份ELISA阳性样本中,检测出其中12份,两者的检测符合率为85.7%。结论初步构建一种登革热综合性蛋白质芯片,结合SPR技术可并行检测多个项目,可为登革热的快速检测提供一个有力的技术平台。

Objective To establish a label-free biosensor for rapid detection of dengue multiplex biomarkers in serum samples, so as to provide a method for the serological diagnosis of dengue virus infection. Methods We employed a surface initiated polymerization strategy to build a polyethylene glycol（PEG）-based surface coating, and followed by the immobilization of Ig M and Ig G antigens and anti NS1 to the surface. The surface plasmon resonance（SPR） measurements were then used to detect the specific dengue biomarkers in sera. Results A total of 262 samples of suspected dengue fever cases were tested and the positive rate was 6.9%（18/262） by the SPR method, which was higher than that by PCR（2.3%, 6/262） and by ELISA（5.3%, 14/262）. Among the 6 positive samples by PCR, NS1 was detected in all those samples by the SPR mehtod and among 14 positive samples by ELISA, NS1 was detected in 12 samples by the SPR mehtod, and the coincidence rate of the ELISA and SPR methods was 85.7%. Conclusion These data suggest that SPR combined with protein array can be used to detect NS1, IgM and IgG simultaneously, which would be potentially useful for dengue fever diagnosis in both endemic and non-endemic areas.