利用表面离子共振系统检测三种禽流感病毒 亚型的研究

Application of the surface plasmon resonance system for identifying subtype of avian influenza A virus

中文关键词:  禽流感病毒 亚型 表面离子共振系统

英文关键词:avian influenza virus  subtype  surface plasmon resonance

中文摘要:

      目的 利用表面离子共振系统建立检测H5、H7、H9三种亚型禽流感病毒的方法。方法 取原核表达的H5、H7、H9亚型的血凝素HA蛋白偶联于CM5芯片上, 利用HBS缓冲液梯度稀释病毒参考抗原并分别与固定浓度的三种亚型的禽流感抗血清进行反应, 采用竞争法制备标准曲线并用以计算样品的浓度和确立检测灵敏度低限。对不同亚型之间及五种其他常见禽类病毒抗体进行特异性检测试验。结果 建立的表面离子共振方法在检测各亚型时相互之间无交叉反应, 未检测到其他禽源病毒的阳性响应信号, HA重组蛋白的检测极限可达0.5 μg/mL。结论 研究表明该方法可用于同步鉴别主要流行的禽流感病毒亚型。

英文摘要:

      Objective To establish a method for detecting avian influenza virus of H5, H7 and H9 subtype using surface plasmon resonance system. Methods The hemagglutinin protein of H5, H7 and H9 subtype which expressed by prokaryotic was coupled on the CM5 chip. The reference antigen of 3 subtypes was diluted gradiently using the HBS buffer and reacted with fixed concentration of 3 subtypes of AIV antiserum respectively. With the preparation of standard curve using competing test, we could calculate the concentration of the sample and detection sensitivity limit. Then the antibody of different subtypes and 5 other poultry disease viruses were tested specificity. Results Various subtypes had no cross reaction between each other, and no positive response of the signals between other poultry virus based on the surface plasmon resonance method. The limit of HA recombinant proteins detection could be reached 0.5 μg/mL. Conclusion The study showed that the method could be used for identifying major epidemic subtypes of avian influenza virus synchronously.