HPV16/18假病毒中和抗体检测方法的验证

目的应用假病毒中和法建立检测血清HPV16/18中和抗体滴度检测方法并进行验证。方法分别采用不同批次假病毒以及不同代次细胞对不同滴度的HPV16/18阳性血清进行多次平行检测,考察这些因素对检验结果的影响;同时通过对抗HPV16/18双价阳性血清、抗HPV16单价阳性血清和抗HPV18单价阳性血清的检测进一步评估中和抗体检测法的准确性、特异性及重复性。结果不同批次假病毒和不同代次细胞对检验结果的影响均在4倍范围内,此外该检测法的准确性、特异性、重复性均在可接受标准范围之内。结论建立的假病毒法可满足中和抗体效价检测的要求,可用于评价疫苗的免疫效果。

Validation of pseudovirion neutralization antibody assay for detecting HPV 16 or HPV 18 antibodies

Objective To develop and validate a pseudovirion-based neutralization assay(PBNS)in determination the titers of serum neutralizing antibodies,including HPV 16 and HPV 18 antibodies.Methods PBNS assay was repeatedly used in detection the titers of both antibodies of HPV 16 and HPV 18 in three different serum samples for observing the effects of different batches of pseudovirion and different passages of cells on the experiment results.The developed method was also evaluated for accuracy,specificity and reproducibility based on the detection antibodies of HPV 16 and HPV 18 by positive sera in both positive(HPV 16 and HPV 18) valent or single positive(either HPV16 or HPV18) valent.Results The effects of different conditions including batches of pseudovirion and passages of cells on the antibody titers varied within a quadruple range.Furthermore,the accuracy,specificity and reproducibility of the developed method were acceptable.Conclusion The established PBNS can meet the detection requirements for neutralizing antibodies of HPV 16 and HPV 18,which could be used in evaluating the immune effect of the related vaccine.