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.

中文翻译：

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加纳阿克拉的HIV-1血清阳性自愿献血者中传播的耐药性突变和亚型多样性。

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方案并确保HIV-1患者中抗逆转录病毒疗法（ART）有效治疗效果的公共卫生策略。在全球范围内，献血者中有关TDR和SD的数据有限。在这项研究中，对加纳阿克拉HIV-1血清阳性献血者的耐药性突变（DRMs）和SD进行了表征。目的采样方法用于从南部地区血液中心收集81份HIV血清阳性血样，并被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），然后通过巢式PCR用特异性引物扩增。预期产品已通过验证，纯化并测序。使用分子进化遗传分析版本6.0（MEGA 6.0），利用RT序列生成具有木村2参数距离的邻居树。在81份血浆样本中，有60份（74％）被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。剩下的21个样本（占26％）被确认为HIV血清阴性。在60个已确认的阳性样本中，分别在RT和PR基因中成功扩增了（32）53％和（28）47％。扩增样品的核苷酸测序表明，两（2）个样品中存在主要的耐药性突变。一个样品中的E138A，另一个使用K65R。发现了HIV-1亚型，包括亚型A，B，CRF02_AG和CRF09_cpx。这项研究发现了主要的耐药性突变，RT基因中的E138A和K65R分别赋予大多数NNRTI和NRTI高水平的抗性。CRF02_AG是最主要的，记录的B型亚型百分比和通过系统发育分析推断的进化关系可能表明可能输入了亚型。但是，需要更前瞻性和详细的分析来确定这种现象。获得的数据将为选择用于ART的药物提供信息，以最大程度地增加加纳的无毒品HIV-1患者的治疗选择。