Reproducibility Assessment of Enzyme-Linked Immunosorbent Assays to Detect Anti-HPV16 L1-Specific IgG1, IgG3, IgA, and IgM Antibodies

酶联免疫吸附试验检测抗 HPV16 L1 特异性 IgG1、IgG3、IgA 和 IgM 抗体的可重复性评估

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作者:Ken Matsui, Heidi Anne Hempel, Gloriana Shelton, Rebecca Ocampo, Troy J Kemp, Yuanji Pan, Ligia A Pinto

Conclusions

Our data showed that these ELISAs are reproducible and detect isotype antibodies to HPV16 L1 across a range of concentrations in 2vHPV-vaccinated participants.

Methods

Seventy-two serum samples collected from participants in the Costa Rica HPV Vaccine Trial (CVT) and immunized with bivalent HPV vaccine (2vHPV) were used for reproducibility assessment. IgG2 and IgG4 levels were too low to be detected. Levels of IgG1, IgG3, IgA, and IgM were measured, and the data were used to calculate intraclass correlation coefficients (ICCs) and coefficients of variation (CVs).

Results

CVs were assessed between technicians (12.8-22.7%) and across days (6.2-30.6%). The overall CVs ranged from 7.7-31.1%. IgM ELISA showed higher CVs (15.8-31.1%) than IgG1, IgG3, and IgA (6.2-22.7%). All ICC values were >98.7%. IgG3 was detected in all samples, while IgG1 and IgA had >86.3% detectability and IgM had 62.1% detectability. Pearson correlational analyses between different antibodies all showed significant correlations (p ≤ 0.001), except when comparing IgGs or IgA to IgM (p = 0.29-0.53). Conclusions: Our data showed that these ELISAs are reproducible and detect isotype antibodies to HPV16 L1 across a range of concentrations in 2vHPV-vaccinated participants.

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