Macchi Beatrice, Frezza Caterina, Marino-Merlo Francesca, Minutolo Antonella, Stefanizzi Valeria, Balestrieri Emanuela, Cerva Carlotta, Sarmati Loredana, Andreoni Massimo, Grelli Sandro, Mastino Antonio
Department of Chemical Science and Technology, University of Rome "Tor Vergata", 00133 Rome, Italy.
Department of Systems Medicine, University of Rome "Tor Vergata", 00133 Rome, Italy.
Pathogens. 2020 Dec 13;9(12):1047. doi: 10.3390/pathogens9121047.
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.
通过聚合酶链反应（PCR）检测血浆中的HIV-1 RNA已被普遍接受为确诊HIV-1感染及监测接受治疗患者病毒载量的最终标准。然而，在某些情况下，该检测方法可能会低估或高估循环病毒或潜伏病毒的复制能力。在本研究中，我们通过一种用于HIV-1患者状态功能筛查的新检测方法对HIV-1逆转录酶（RT）活性进行了评估。为此，我们首次在血液样本上采用了一种实时RT定量PCR检测方法的改良版本，该方法用于评估化合物的HIV-1-RT抑制活性。该研究分析了28例HIV-1感染患者的血液样本，这些患者的病毒血症和免疫值范围广泛。结果表明，以所评估检测方法获得的循环阈值表示的血浆HIV-1 RT水平与患者的血浆HIV-1-RNA水平呈高度显著负相关。因此，我们在本研究中创建了一种HIV-1 RT定量PCR检测方法，它可被视为一种有前景的基础，用于开发一种能够提供HIV-1感染患者功能病毒学状态信息的额外工具。
