Accuracy of real-time PCR and digital PCR for the monitoring of total HIV DNA under prolonged antiretroviral therapy

Constance Renault (1, 2, 3) , Karine Bolloré (2, 3, 1) , Amandine Pisoni (2, 3, 1, 4) , Camille Motto-Ros (2, 1, 3) , Philippe van de Perre (2, 3, 1, 4) , Jacques Reynes (5, 4) , Edouard Tuaillon (2, 4, 3, 1)

1 UA - Université des Antilles
2 PCCEI - Pathogenesis and Control of Chronic and Emerging Infections
3 Etablissement français du don du sang [Montpellier]
4 CHRU Montpellier - Centre Hospitalier Régional Universitaire [Montpellier]
5 TransVIHMI - Recherches Translationnelles sur le VIH et les maladies infectieuses endémiques et émergentes

Total HIV DNA is a standard marker to monitor the HIV reservoir in people living with HIV. We investigated HIV DNA quantification accuracy by a real-time PCR kit (qPCR) and digital PCR (dPCR) method within the same set of primers and probes. Among 48 aviremic patients followed for up to 7 years with qPCR, the mean coefficient of variation of total HIV DNA between two successive measurements was 77% (± 0.42log 10 HIVDNA copies/10 6 PBMC). The total HIV DNA quantified by the two PCR methods has a high correlation (0.99 and 0.83, for 8E5 and PLHIV samples, respectively), but we observed better repeatability and reproducibility of the dPCR compared to the qPCR (CV of 11.9% vs. 24.7% for qPCR, p-value = 0.024). Furthermore, we highlighted a decay of the number of HIV copies in the 8E5 cell line qPCR standard over time (from 0.73 to 0.43 copies per cell), contributing to variations of HIV DNA results in patients whose HIV reservoir should be theoretically stabilized. Our study highlighted that absolute quantification of total HIV DNA by dPCR allows more accurate monitoring of the HIV reservoir than qPCR in patients under prolonged antiretroviral therapy.

Domaines: Sciences du Vivant [q-bio]; Médecine humaine et pathologie

Constance Renault, Karine Bolloré, Amandine Pisoni, Camille Motto-Ros, Philippe van de Perre, et al.. Accuracy of real-time PCR and digital PCR for the monitoring of total HIV DNA under prolonged antiretroviral therapy. Scientific Reports, 2022, 12 (1), pp.9323. ⟨10.1038/s41598-022-13581-8⟩. ⟨hal-03758868⟩