Evaluation of the performance of a qPCR-based assay for HIV-1 viral load determination.

Objective: According to the YY/T 1182-2010 standard of the People's Republic of China on nucleic acid amplification test reagents (kits) for medical industry, the accuracy, precision, linear range, and analytic sensitivity of HIV-1 standardized quality control products should be assessed. The Geneway HIV-1 Nucleic Acid Detection Kit from China has been successfully registered with the National Medical Products Administration. Here, we aimed to assess for the first time its detection performance.

Methods: The accuracy, precision, analytic sensitivity, and linearity of the Geneway HIV-1 nucleic acid quantification test kit were analyzed using a series of diluted standard control samples of HIV-1 negative plasma. Clinical plasma samples were collected from 163 HIV-infected patients and 38 HIV-negative patients. The detection performance of the Geneway assay was compared with that of the US FDA-approved COBAS AmpliPrep/COBAS® Taqman® HIV-1 test (Roche), version 2.0, for viral load (VL) monitoring.

Results: The absolute deviation of the assay between the logarithm of the measured concentration and the logarithm of the expected concentration did not exceed ±0.5 logarithmic units. All coefficients of variation (CV%) for the assays were within 5%, indicating good precision in the detection. The linearity of quantitation was excellent (r = 0.999). Overall agreement was observed in 198 of the 201 specimens (98.51%), with a kappa value of 0.953. Bland-Altman analysis revealed an average difference of 0.030 between the two assays, with 95.95% (142/148) of the differences falling within the 95% confidence limits of agreement (-0.50, 0.56). Linear regression results demonstrated a strong linear correlation between the two assays, with a high Pearson correlation coefficient (r = 0.980) and coefficient of determination (R2 = 0.960, p < 0.001).

Conclusions: The Geneway HIV-1 VL assay demonstrated excellent accuracy, precision, analytic sensitivity, and linearity. Compared to the Roche assay, the Geneway assay showed good performance for HIV-1 VL detection, supporting its use in clinical practice.