A method to alleviate false-positive results of the Elecsys HIV combi PT assay

Xiaolan Lu, Minghong Zhang, Wen Liu, Nan Sheng, Qin Du, Maoxin Zhang, Xiaolan Guo, Guangrong Wang, Qiang Wang, Xiaolan Lu, Minghong Zhang, Wen Liu, Nan Sheng, Qin Du, Maoxin Zhang, Xiaolan Guo, Guangrong Wang, Qiang Wang

Abstract

To explore the effects of urea dissociation on reducing false-positive results of the Elecsys HIV combi PT assay. A retrospective analysis was used to evaluate the false-positive rate of the Elecsys HIV combi PT assay. Six false-positive sera, six positive sera and six sera from patients with early HIV infection were collected. Dissociation was performed using 1 mol/L, 2 mol/L, 4 mol/L, 6 mol/L, or 8 mol/L urea, and HIV screening assay were then detected to select the appropriate concentration of urea dissociation. Next, 55 false-positive sera and 15 sera from early HIV infection were used to verify the best concentration of urea to achieve dissociation. Retrospective analysis showed that the COI of the Elecsys HIV combi PT assay in false-positive sera ranged from 1.0 to 200.0, and approximately 97.01%(227/234) of false-positive sera were in the range of 1.0-15.0. The avidity index (AI) in positive and false-positive sera decreased as the urea dissociation concentration increased. When the dissociation concentration was 6 mol/L, the AI of false-positive serum was between 0.0234 and 0.2567, and the AI of early HIV infection sera was between 0.4325 and 0.5017. The difference in AI between false-positive and positive samples was significant. When negativity was defined as an AI of less than 0.3970, the sensitivity and specificity were 100.0% and 100.0%, respectively. Urea-mediated dissociation could significantly reduce the false-positive rate of the Elecsys HIV combi PT assay with a low COI. Our findings provided a reference for distinguishing positive and false-positive of the Elecsys HIV combi PT assay.

Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Distribution of screening values of COIs in false-positive and positive specimens.
Figure 2
Figure 2
First western blot analysis of human sera for urea dissociation tests. S standard line. Lanes 1–6: sera obtained from patients with false-positive results, showing no bands or only the P24 band for specific antigens of HIV-1. Lanes 7–12: sera obtained from patients with early HIV infection, showing no bands or only one distinct band for specific antigens of HIV-1. Lanes 13–18: sera obtained from patients with positive results, showing more than two distinct bands for specific antigens of HIV-1.
Figure 3
Figure 3
Western blot analysis of false-positive results and early HIV infections after follow-up for 2 weeks to 3 months. S standard line. Lanes 1–6: sera obtained from patients with false-positive results, showing no bands or only a P24 band for specific antigens of HIV-1. Lanes 7–12: sera obtained from patients with early HIV infection, showing more than two distinct bands for specific antigens of HIV-1.
Figure 4
Figure 4
Urea dissociation test to determine the appropriate dissociation concentration.
Figure 5
Figure 5
Validation experiment for 6 mol/L urea. After dissociation with 6 mol/L urea, the AI of false-positive serum samples was measured, and the sensitivity and specificity values were determined.
Figure 6
Figure 6
Flowchart depicting the study protocol.

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