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采用液滴数字 PCR 技术评价 HIV 感染长期持续病毒学应答患者的丙型肝炎病毒 RNA 持续性。

Evaluation of hepatitis C viral RNA persistence in HIV-infected patients with long-term sustained virological response by droplet digital PCR.

机构信息

Unidad de Enfermedades Infecciosas. Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC), Hospital Universitario Reina Sofía de Córdoba, Universidad de Córdoba, Córdoba, Spain.

Unidad Gestión Clínica Enfermedades Infecciosas, Hospital La Línea, AGS Campo de Gibraltar, Cádiz, Spain.

出版信息

Sci Rep. 2019 Aug 29;9(1):12507. doi: 10.1038/s41598-019-48966-9.

Abstract

Several studies have reported the persistence of HCV RNA in liver and/or peripheral blood mononuclear cells (PBMCs) in spite of undetectable viremia in patients who have achieved sustained virological response (SVR). This event, defined as occult HCV infection, remains controversial and low titers of persistent virus may be underestimated because it has not yet been analyzed by a highly sensitive test such as droplet digital PCR (ddPCR). This method provides an alternate ultra-sensitive detection technique for very low numbers of copies of viral RNA or DNA. The aim of this study was to evaluate the persistence of HCV in HIV-coinfected patients with long-term SVR using ddPCR. For each patient, the presence of HCV RNA in serum and PBMCs at baseline was determined by nested RT-ddPCR. Patients with HCV RNA in PBMCs at baseline were followed until the end of the study. One hundred and twenty-three patients were analyzed for persistence of HCV RNA in serum and PBMCs. Persistence of HCV was not found in serum in any patient. HCV RNA was detected in PBMCs in one patient (0.81%; 95% CI: 0.04-3.94) and resolved spontaneously during follow-up. Persistence of HCV RNA in PBMCs is not a common event in HIV/HCV co-infected patients with long-term SVR evaluated by RT-ddPCR.

摘要

尽管达到持续病毒学应答 (SVR) 的患者血液中无法检测到病毒载量,但多项研究报告称,其肝组织和/或外周血单个核细胞 (PBMC) 中仍存在 HCV RNA。这种现象被定义为隐匿性 HCV 感染,目前仍存在争议,因为尚未通过高灵敏度检测方法(如液滴数字 PCR (ddPCR))进行分析,因此可能低估了低水平持续性病毒的存在。该方法为病毒 RNA 或 DNA 的极低拷贝数提供了一种替代的超灵敏检测技术。本研究旨在使用 ddPCR 评估 HIV 合并感染且长期 SVR 的患者中 HCV 的持续性。对于每位患者,均通过巢式 RT-ddPCR 检测基线时血清和 PBMC 中 HCV RNA 的存在情况。基线时 PBMC 中有 HCV RNA 的患者会一直随访至研究结束。对 123 例患者进行了血清和 PBMC 中 HCV RNA 持续性的分析。未在任何患者的血清中发现 HCV 持续性。在一名患者(0.81%;95%CI:0.04-3.94)的 PBMC 中检测到 HCV RNA,且在随访期间自行消退。通过 RT-ddPCR 评估,HIV/HCV 合并感染且长期 SVR 的患者中,PBMC 中 HCV RNA 的持续性并不常见。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e95/6715682/b526647732e4/41598_2019_48966_Fig1_HTML.jpg

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