H1N1亚型流感假病毒评价中和抗体的研究

The study on neutralizing antibody detection with H1N1 subtype pseudotyped influenza viral particles

目的 采用流感假病毒检测高致病性流感病毒中和抗体,以评价人群保护力.方法 构建H1N1亚型流感假病毒作为病毒抗原,同时采用血抑试验和细胞病变中和试验方法,检测NIBSC标准血清、单克隆抗体（McAb）、羊抗血清以及人群样本血清中的抗体滴度,并与天然完整病毒相比较.结果 在两种检测方法中,假病毒与天然完整病毒检测McAb、NIBSC标准血清和羊抗血清的抗体滴度结果接近,差异无统计学意义（P＞0.05）.检测采自2013年的300份人群血清H1N1亚型流感病毒抗体的阳性率,人群中假病毒与天然完整病毒检测结果呈显著的高度正相关关系（P＜0.001）,相关系数为0.96.人群中抗H1N1抗体阳性率达54.7％,抗体滴度25.70 （22.39,29.51）.结论 假病毒可用于流感中和抗体的评价.

Objective To evaluate the effectiveness of pseudotyped influenza viral particles （pp） in neutralizing antibody （nAb） detection.Methods Pseudotyped particles of A （H1 N 1） pmd09 （A/ California/7/2009） were generated.Pp in nAb detection were compared concurrently with the corresponding viruse by a hemagglutination inhibition test,as well as cytopathic effect-（CPE） and fluorescence-based neutralization assays.Results No significant difference between pp and the corresponding virus was observed in nAb detection of anti-H1 N1 monoclonal antibody,NIBSC antiserum and goat immune serum by two assays （P 〉0.05）.Moreover,pp and its corresponding virus had significant positive correlation （P 〈 0.001）,with the correlation coefficient near 1 in population sera detection of 300 samples collected in 2013.The positive rate of H1 N1 antibody was 54.7% in population sera detection,and the geometric mean titers （95 % confidence interval） [GMT （95 % CI）] was 25.70 （22.39,29.51）.Conclusion Pp was reliable in neutralizing antibody detection.