发热伴血小板减少综合征布尼亚病毒抗原定量检测方法的建立

Establishment of a quantitative detection method for SFISV antigen

目的建立定量检测发热伴血小板减少综合征布尼亚病毒（severe fever with thrombocytopenia syndrome bunyavirus，SFTSV）抗原的方法，以用于疫苗生产过程中sFrSV抗原含量的检测。方法用SFTSV抗原免疫BALB／c小鼠，制备抗SFTSV单克隆抗体。抗体纯化后以辣根过氧化物酶进行标记，建立双抗体夹心ELISA法。确定该法的线性范围和灵敏度，并对该法的准确性、精密性、专属性进行验证。结果建立的ELISA法的线性范围为0．117～2．000mg／L，定量限为0．117mg／L，线性的决定系数为0．9903。该法具有良好的准确性和精密度，样品回收率为98％，变异系数均〈8％。该法的专属性良好，对6株不同sFTSV的检测结果均为阳性，而与其他布尼亚病毒、Veto细胞培养上清及其他生产辅料均无交叉反应。结论成功建立了SFTSV抗原的定量检测方法，为疫苗生产的质量控制奠定了基础。

Objective To establish a quantitative detection method for severe fever with thrombocytopenia syndrome bunyavirus （SFTSV） antigen to detect contents of SFTSV antigen during SFTSV vaccine production. Methods Monoclonal antibody was prepared by immunizing BALB/c mouse with SFTSV antigen, and then purified and labeled with horseradish peroxidase to establish a double antibody sandwich ELISA method. Linear range and sensitivity of the ELISA method were detected, and accuracy, precision and specificity of the ELISA method were validated. Results Linear range, quantification limit and determination coefficient of the ELISA method were 0. 117-2. 000 mg/L, 0. 117 mg/L and 0. 990 3, respectively. Accuracy and precision of the ELISA method were good. Recoveries of internal reference were 98% and variation coefficient were all lower than 8%. The ELISA method had good specificity. The results for detecting 6 strains of SFTSV with the ELISA method were all positive, and no reactions were observed with other bunyaviruses, Vero cells and other excipients. Conclusion A quantitative detection method for SFTSV antigen is successfully established and lays the foundation for quality control during SFTSV vaccine production.