Department of Virology, Noguchi Memorial Institute for Medical Research (NMIMR), University of Ghana, Accra, Ghana.
Department of Medical Microbiology, School of Medicine and Dentistry, University of Ghana, Accra, Ghana.
Virol J. 2020 Jul 24;17(1):114. doi: 10.1186/s12985-020-01386-y.
Detection of HIV-1 transmitted drug resistance (TDR) and subtype diversity (SD) are public health strategies to assess current HIV-1 regimen and ensure effective therapeutic outcomes of antiretroviral therapy (ART) among HIV-1 patients. Globally, limited data exist on TDR and SD among blood donors. In this study, drug resistance mutations (DRMs) and SD amongst HIV-1 sero-positive blood donors in Accra, Ghana were characterized.
Purposive sampling method was used to collect 81 HIV sero-positive blood samples from the Southern Area Blood Center and confirmed by INNO-LIA as HIV-1 and/or HIV-2. Viral RNA was only extracted from plasma samples confirmed as HIV-1 positive. Complementary DNA (cDNA) was synthesized using the RNA as a template and subsequently amplified by nested PCR with specific primers. The expected products were verified, purified and sequenced. Neighbour-joining tree with the Kimura's 2-parameter distances was generated with the RT sequences using Molecular Evolutionary Genetic Analysis version 6.0 (MEGA 6.0).
Out of the 81 plasma samples, 60 (74%) were confirmed as HIV-1 sero-positive by INNO-LIA HIVI/II Score kit with no HIV-2 and dual HIV-1/2 infections. The remaining samples, 21 (26%) were confirmed as HIV sero-negative. Of the 60 confirmed positive samples, (32) 53% and (28) 47% were successfully amplified in the RT and PR genes respectively. Nucleotide sequencing of amplified samples revealed the presence of major drug resistance mutations in two (2) samples; E138A in one sample and another with K65R. HIV-1 Subtypes including subtypes A, B, CRF02_AG and CRF09_cpx were found.
This study found major drug resistance mutations, E138A and K65R in the RT gene that confer high level resistance to most NNRTIs and NRTI respectively. CRF02_AG was most predominant, the recorded percentage of subtype B and the evolutionary relationship inferred by phylogenetic analysis may suggest possible subtype importation. However, a more prospective and detailed analysis is needed to establish this phenomenon. The data obtained would inform the selection of drugs for ART initiation to maximize therapeutic options in drug-naïve HIV-1 patients in Ghana.
检测 HIV-1 传播的耐药性（TDR）和亚型多样性（SD）是评估当前 HIV-1 治疗方案并确保 HIV-1 患者接受有效的抗病毒治疗（ART）的公共卫生策略。在全球范围内，有关血液供者中 TDR 和 SD 的数据有限。在这项研究中，我们对加纳阿克拉的 HIV-1 血清阳性血液供者中的耐药突变（DRMs）和 SD 进行了描述。
采用目的抽样法收集来自南部地区血液中心的 81 份 HIV 血清阳性血液样本，并通过 INNO-LIA 确认为 HIV-1 和/或 HIV-2。仅从确认为 HIV-1 阳性的血浆样本中提取病毒 RNA。使用 RNA 作为模板合成互补 DNA（cDNA），然后使用巢式 PCR 和特异性引物进行扩增。验证、纯化和测序预期产物。使用分子进化遗传分析版本 6.0（MEGA 6.0），根据 RT 序列生成基于 Kimura 2-参数距离的邻接法树。
在 81 个血浆样本中，60 个（74%）通过 INNO-LIA HIVI/II 评分试剂盒确认为 HIV-1 血清阳性，无 HIV-2 和双重 HIV-1/2 感染。其余样本，21 个（26%）被确认为 HIV 血清阴性。在 60 个确认为阳性的样本中，RT 基因分别成功扩增了（32）53%和（28）47%，PR 基因分别成功扩增了（32）53%和（28）47%。扩增样本的核苷酸测序显示，在两个样本中存在主要耐药突变；一个样本中的 E138A 和另一个样本中的 K65R。发现了 HIV-1 亚型，包括亚型 A、B、CRF02_AG 和 CRF09_cpx。
本研究在 RT 基因中发现了主要的耐药突变 E138A 和 K65R，它们分别对大多数 NNRTIs 和 NRTI 产生高水平耐药。CRF02_AG 最为普遍，记录的 B 亚型百分比和系统发育分析推断的进化关系可能表明存在亚型输入。然而，需要进行更前瞻性和详细的分析来确定这一现象。获得的数据将为加纳开始抗逆转录病毒治疗的药物选择提供信息，以最大限度地扩大初治 HIV-1 患者的治疗选择。
