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.

Macchi, B., Frezza, C., Marino-Merlo, F., Minutolo, A., Stefanizzi, V., Balestrieri, E., et al. (2020). Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients. PATHOGENS, 9(12), 1-9 [10.3390/pathogens9121047].

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

Macchi, Beatrice;Frezza, Caterina;Minutolo, Antonella;Balestrieri, Emanuela;Sarmati, Loredana;Andreoni, Massimo;Grelli, Sandro;
2020-12-13

Abstract

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.
13-dic-2020
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/19 - MICROBIOLOGIA GENERALE
Settore CHIM/08 - CHIMICA FARMACEUTICA
Settore MED/17 - MALATTIE INFETTIVE
English
Con Impact Factor ISI
HIV diagnostics
human immunodeficiency virus
quantitative PCR assay
reverse transcriptase
viral load
Macchi, B., Frezza, C., Marino-Merlo, F., Minutolo, A., Stefanizzi, V., Balestrieri, E., et al. (2020). Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients. PATHOGENS, 9(12), 1-9 [10.3390/pathogens9121047].
Macchi, B; Frezza, C; Marino-Merlo, F; Minutolo, A; Stefanizzi, V; Balestrieri, E; Cerva, C; Sarmati, L; Andreoni, M; Grelli, S; Mastino, A
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/262815
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