Deletsu Selase D, Maina Edward K, Quaye Osbourne, Ampofo William K, Awandare Gordon A, Bonney Evelyn Y
West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology.
Department of Virology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon-Accra, Ghana.
Medicine (Baltimore). 2020 Feb;99(7):e18777. doi: 10.1097/MD.0000000000018777.
This study sought to determine the dominant circulating human immunodeficiency virus type 1 (HIV-1) subtype and associated drug resistance mutations in Ghana.This cross-sectional study was conducted with archived samples collected from patients who received care at 2 hospitals in Ghana from 2014 to 2016. Blood samples were earlier processed into plasma and peripheral blood mononuclear cells and stored at -80 °C. Ribonucleic acid (RNA) was extracted from the archived plasma. Two HIV-1 genes; protease and reverse transcriptase, were amplified, sequenced using gene-specific primers and analyzed for subtype and drug resistance mutations using the Stanford HIV Database.Of 16 patient samples successfully sequenced, we identified the predominance of HIV-1 subtype CRF02_AG (11/16, 68%). Subtypes G (2/16, 13%), dual CRF02_AG/G (2/16, 13%), and CRF01_AE (1/16, 6%) were also observed. Major nucleoside reverse transcriptase inhibitor (NRTI) resistance mutations, M184I/V, D67N, T215F, and K70R/E were found. Non-nucleoside reverse transcriptase inhibitor (NNRTI) resistance mutations, K103N, Y181C, V90I, F227L, and V106A were also prevalent. Additionally, and at a lower level, protease inhibitor (PI)-resistance mutations, M46I, I54 V, V82A, L90 M, and I471 V, were also present in the sequences from antiretroviral therapy (ART)-experienced individuals. Two NRTI-associated drug resistance mutations (DRMs) (D67N and T69N) were present in sequences from 1 ART-naive individual.HIV-1 subtype CRF02_AG was most frequently detected in this study thus confirming earlier reports of dominance of this subtype in the West-African sub-region and Ghana in particular. The detection of these drug resistance mutations in individuals on first-line regimen composed of NRTI and NNRTI is an indication of prolonged drug exposure without viral load monitoring. Routine viral load monitoring is necessary for early detection of virologic failure and drug resistance testing will inform appropriate choice of regimens for such patients.
本研究旨在确定加纳主要的循环1型人类免疫缺陷病毒（HIV-1）亚型及相关耐药突变。这项横断面研究使用的是2014年至2016年期间从加纳两家医院接受治疗的患者收集的存档样本。血液样本早前已处理成血浆和外周血单核细胞，并储存在-80°C。从存档血浆中提取核糖核酸（RNA）。使用基因特异性引物扩增并测序两个HIV-1基因：蛋白酶基因和逆转录酶基因，并使用斯坦福HIV数据库分析亚型和耐药突变。在成功测序的16份患者样本中，我们确定HIV-1 CRF02_AG亚型占主导地位（11/16，68%）。还观察到G亚型（2/16，13%）、双重CRF02_AG/G亚型（2/16，13%）和CRF01_AE亚型（1/16，6%）。发现了主要的核苷类逆转录酶抑制剂（NRTI）耐药突变，如M184I/V, D67N, T215F和K70R/E。非核苷类逆转录酶抑制剂（NNRTI）耐药突变，如K103N, Y181C, V90I, F227L和V1也很普遍。此外，在接受抗逆转录病毒治疗（ART）的个体序列中，还存在较低水平的蛋白酶抑制剂（PI）耐药突变，如M46I, I54V, V82A, L90M和I471V。在1例初治个体的序列中存在两个与NRTI相关的耐药突变（DRMs）（D67N和T69N）。在本研究中，HIV-1 CRF02_AG亚型最常被检测到，从而证实了早期报告中该亚型在西非次区域尤其是加纳占主导地位的情况。在由NRTI和NNRTI组成的一线治疗方案的个体中检测到这些耐药突变，表明在没有病毒载量监测的情况下长期用药。常规病毒载量监测对于早期发现病毒学失败是必要的，而耐药性检测将为这类患者选择合适的治疗方案提供依据。
