Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients

• Main Authors: Beatrice Macchi, Caterina Frezza, Francesca Marino-Merlo, Antonella Minutolo, Valeria Stefanizzi, Emanuela Balestrieri, Carlotta Cerva, Loredana Sarmati, Massimo Andreoni, Sandro Grelli, Antonio Mastino
• Format: Article
• Language: English
• Published: MDPI AG 2020-12-01
• Series: Pathogens
• Subjects: human immunodeficiency virus; HIV diagnostics; viral load; reverse transcriptase; quantitative PCR assay
• Online Access: https://www.mdpi.com/2076-0817/9/12/1047

Testing HIV-1 RNA in plasma by PCR is universally accepted as the ultimate standard to confirm diagnosis of HIV-1 infection and to monitor viral load in patients under treatment. However, in some cases, this assay could either underestimate or overestimate the replication capacity of a circulating or latent virus. In the present study, we performed the assessment of evaluating the HIV-1 reverse transcriptase (RT) activity by means of a new assay for the functional screening of the status of HIV-1 patients. To this purpose, we utilized, for the first time on blood samples, an adapted version of a real-time RT quantitative PCR assay, utilized to evaluate the HIV-1-RT inhibitory activity of compounds. The study analyzed blood samples from 28 HIV-1-infected patients, exhibiting a wide range of viremia and immunological values. Results demonstrated that plasma HIV-1 RT levels, expressed as cycle threshold values obtained with the assay under appraisal, were inversely and highly significantly correlated with the plasma HIV-1-RNA levels of the patients. Thus, an HIV-1 RT quantitative PCR assay was created which we describe in this study, and it may be considered as a promising basis for an additional tool capable of furnishing information on the functional virological status of HIV-1-infected patients.