Abstract
INTRODUCTION: A novel recombinant antigen-based capture enzyme immunoassay (RAg-CEIA) was optimized and used to determine technical parameters for estimating human immunodeficiency virus type 1 (HIV-1) incidence in China. METHODS: We employed orthogonal experimental design to optimize RAg-CEIA by adjusting raw material dilution ratios. The assay was used to measure normalized optical density (ODn) values in 171 longitudinal plasma specimens from 51 HIV-1 seroconverting individuals, plotted against estimated days post-seroconversion. We determined the optimal ODn threshold value for differentiating recent from long-term infections and calculated the mean duration of recent infection (MDRI) for incidence estimation. The false recent rate (FRR) was determined using 481 HIV-1 antibody-positive specimens with infection durations exceeding twice the MDRI. RESULTS: Optimal RAg-CEIA parameters were established with a raw material dilution ratio of 1/12 for calibrator preparation and an enzyme conjugate titer of 1:1200. ODn values demonstrated consistent temporal increases across HIV-1 seroconverting individuals, though with notable kinetic heterogeneity in individual responses. The optimal ODn threshold value of 0.8 for distinguishing recent from long-term infections corresponded to an MDRI of 205 days and an FRR of 4.78%. CONCLUSIONS: The optimized RAg-CEIA effectively differentiates recent from long-term HIV-1 infections at the population level, enabling reliable HIV-1 incidence estimation in China.