Development of an in-house ELISA to detect anti-HPV16-L1 antibodies in serum and dried blood spots
Development of an in-house ELISA to detect anti-HPV16-L1 antibodies in serum and dried blood spots
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DOI：
10.1016/j.jviromet.2018.10.007
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发表时间：
2019-02-01
影响因子：
3.1
通讯作者：
Cuschieri, Kate
中科院分区：
文献类型：
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作者：
Bhatia, Ramya;Stewart, June;Cuschieri, Kate
测量抗hpv抗体水平对于监测对自然感染和疫苗接种的免疫反应非常重要。本研究开发了一种ELISA检测HPV-16L1抗体，并在血清和干血斑中进行了验证。我们开发了一种ELISA检测抗hpv - 16l1抗体的IgA和IgG水平。根据世卫组织国际标准血清对该检测方法进行了标准化，并对苏格兰参加阴道镜检查诊所的妇女的血清、干血斑和宫颈液基细胞学样本进行了验证。同时测定血清样品的抗体亲和力指数。参加常规阴道镜检查的妇女血清中HPV 16-L1特异性IgG和IgA的平均水平分别为7.3单位/ml和8.1单位/ml。血清和干血斑洗脱液中IgG和IgA含量显著相关，表明后者可作为抗体水平的可靠指标。平均IgG贪婪指数为35% (95% CI为25%-45%)，提示既往自然感染的历史挑战。这种酶联免疫吸附试验有可能用于抗体流行病学和实地研究，如果与免疫度测量相结合，可能用于疫苗反应和失败的个案监测。
Measuring anti-HPV antibody levels is important for surveillance of the immunological response to both natural infection and vaccination. Here, an ELISA test for measurement of HPV-16L1 antibodies was developed and validated in sera and dried blood spots.An in-house ELISA was developed for measuring anti-HPV-16L1 IgA and IgG levels. The assay was standardized against WHO international standard serum and validated on serum, dried blood spots and cervical liquid based cytology samples from women attending colposcopy clinics in Scotland. Antibody avidity index was also measured in serum samples.The average HPV 16-L1 specific IgG and IgA levels measured in sera, in women attending a routine colposcopy service were 7.3 units/ml and 8.1 units/ml respectively. Significant correlations between serum and dried blood spot eluates for both IgG and IgA were observed indicating that the latter serve as a credible proxy for antibody levels. Average IgG Avidity Index was 35% (95% CI 25%-45%) suggesting previous, historical challenge with natural infection. This ELISA has potential for use in epidemiological and field studies of antibody prevalence and if coupled with avidity measurement may be of use in individual case monitoring of vaccine responses and failures.