SARS病毒快速检测膜研制

Development of a Dot Immunofiltration Assay for Rapid Identification of (SARS-CoV)

建立一种快速检测严重急性呼吸综合症冠状病毒(SARS-CoV)的方法。从SARS冠状病毒刺突蛋白基因中扩增出羧基端片段,克隆并表达。用经Ni-NTA树脂纯化的表达蛋白作为抗原包被硝酸纤维素膜,捕获感染SARS病人血清中的抗体。抗原抗体通过渗滤在膜上反应,5min肉眼即可观察到结果。通过比较不同试剂用量条件下的结果确定检测抗原的最佳浓度。通过本产品检测已经过ELISA验证的50份血清样品证明两种检测结果无显著性差异(P>0.05)。与ELISA相比,该检测方法特异性、灵敏性几乎相当,但具有简单、快速、成本低廉等ELISA所无法比拟的优点,适合于初步诊断和流行病学调查。

In order to develop a clinical diagnostic tool for rapid detection of SARS-CoV（severe acute respiratory syndrome-associated coronavirus）, a dot immunofihration method was established. The C-terminal portion of the spike protein gene was amplified from the SARS CoV genome, cloned and expressed. The soluble protein purified by Ni-NTA resin and used as antigen was coated on the nitrocellulose membrane to capture the antibody in sera of patients with SARS infection. It was found that the results of this assay could be observed by naked eyes within 5 minutes. Optimal concentration of the antigen was determined by tests of serially diluted antigen reacting with that of antiserum. By using this method to examine 50 serum samples detected by commercial ELISA previously, the two different methods showed no distinct difference （P 〉 0.05）. This method has almost the same specificity and sensitivity when compared with ELISA, and besides, it also has the characteristics of rapidity,simplicity and cheapness which ELISA can not afford, and thus, this method can be used for primary SARS diagnosis and epidemiological screening.