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Updated: Jun 4, 2025
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
Study on the Technical Parameters for Estimating HIV-1 Incidence by Using a Recombinant Antigen-based Capture Enzyme
Wenli Liang1, Jibao Wang2, Hongxia Yan3
1National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
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.
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