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

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 o...

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Autores principales: Macchi, Beatrice, Frezza, Caterina, Marino-Merlo, Francesca, Minutolo, Antonella, Stefanizzi, Valeria, Balestrieri, Emanuela, Cerva, Carlotta, Sarmati, Loredana, Andreoni, Massimo, Grelli, Sandro, Mastino, Antonio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7763350/
https://www.ncbi.nlm.nih.gov/pubmed/33322208
http://dx.doi.org/10.3390/pathogens9121047
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author Macchi, Beatrice
Frezza, Caterina
Marino-Merlo, Francesca
Minutolo, Antonella
Stefanizzi, Valeria
Balestrieri, Emanuela
Cerva, Carlotta
Sarmati, Loredana
Andreoni, Massimo
Grelli, Sandro
Mastino, Antonio
author_facet Macchi, Beatrice
Frezza, Caterina
Marino-Merlo, Francesca
Minutolo, Antonella
Stefanizzi, Valeria
Balestrieri, Emanuela
Cerva, Carlotta
Sarmati, Loredana
Andreoni, Massimo
Grelli, Sandro
Mastino, Antonio
author_sort Macchi, Beatrice
collection PubMed
description 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.
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spelling pubmed-77633502020-12-27 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 Marino-Merlo, Francesca Minutolo, Antonella Stefanizzi, Valeria Balestrieri, Emanuela Cerva, Carlotta Sarmati, Loredana Andreoni, Massimo Grelli, Sandro Mastino, Antonio Pathogens Communication 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. MDPI 2020-12-13 /pmc/articles/PMC7763350/ /pubmed/33322208 http://dx.doi.org/10.3390/pathogens9121047 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Macchi, Beatrice
Frezza, Caterina
Marino-Merlo, Francesca
Minutolo, Antonella
Stefanizzi, Valeria
Balestrieri, Emanuela
Cerva, Carlotta
Sarmati, Loredana
Andreoni, Massimo
Grelli, Sandro
Mastino, Antonio
Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients
title Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients
title_full Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients
title_fullStr Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients
title_full_unstemmed Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients
title_short Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients
title_sort appraisal of a simple and effective rt-qpcr assay for evaluating the reverse transcriptase activity in blood samples from hiv-1 patients
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7763350/
https://www.ncbi.nlm.nih.gov/pubmed/33322208
http://dx.doi.org/10.3390/pathogens9121047
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