Botswana Harvard AIDS Institute Partnership Gaborone, Botswana.
Department of Medical Laboratory Sciences, School of Allied Health Professionals, University of Botswana, Gaborone, Botswana.
PLoS One. 2019 Nov 21;14(11):e0224292. doi: 10.1371/journal.pone.0224292. eCollection 2019.
Roll-out of Integrase Strand Transfer Inhibitors (INSTIs) such as dolutegravir for HIV combination antiretroviral therapy (cART) in sub-Saharan Africa necessitates the development of affordable HIV drug resistance (HIVDR) assays targeting the Integrase gene. We optimised and evaluated an in-house integrase HIV-1 drug resistance assay (IH-Int) and compared it to a commercially available assay, ViroSeq™ Integrase Genotyping kit (VS-Int) amongst HIV-1 clade C infected individuals.
We used 54 plasma samples from treatment naïve participants and one plasma sample from a patient failing INSTI based cART. Specimens were genotyped using both the VS-Int and IH-Int assays. Stanford HIV drug resistance database were used for integrase resistance interpretation. We compared the major and minor resistance mutations, pairwise nucleotide and amino-acid identity, costs and assay time.
Among 55 specimens tested with IH-Int, 53 (96.4%) successfully amplified compared to 45/55 (81.8%) for the VS-Int assay. The mean nucleotide and amino acid similarity from 33 paired sequences was 99.8% (SD ± 0.30) and 99.8% (SD ± 0.39) for the IH-Int and VS-Int assay respectively. The reagent cost/sample were 32 USD and 147 USD for IH-Int and VS-Int assay, respectively. All sequenced samples were confirmed as HIV-1 subtype C.
The IH-Int assay had a high amplification success rate and high concordance with the commercial assay. It is significantly cheaper compared to the commercial assay. Our assay has the needed specifications for routine monitoring of participants on Dolutegravir based regimens in Botswana.
在撒哈拉以南非洲地区推出整合酶链转移抑制剂（INSTIs），如多替拉韦，用于 HIV 联合抗逆转录病毒治疗（cART），需要开发针对整合酶基因的负担得起的 HIV 耐药性（HIVDR）检测方法。我们优化并评估了一种内部整合酶 HIV-1 耐药性检测（IH-Int），并将其与市售的 ViroSeq™ 整合酶基因分型试剂盒（VS-Int）在 HIV-1 型 C 群感染者中进行了比较。
我们使用了 54 份来自初治患者的血浆样本和 1 份来自正在接受 INSTI 为基础的 cART 治疗失败患者的血浆样本。使用 VS-Int 和 IH-Int 两种检测方法对标本进行基因分型。斯坦福 HIV 耐药数据库用于整合酶耐药性解释。我们比较了主要和次要耐药突变、核苷酸和氨基酸的配对差异、成本和检测时间。
在 55 份用 IH-Int 检测的标本中，有 53 份（96.4%）成功扩增，而 VS-Int 检测的成功率为 45/55（81.8%）。33 对配对序列的平均核苷酸和氨基酸相似度分别为 IH-Int 检测的 99.8%（SD ± 0.30）和 99.8%（SD ± 0.39），VS-Int 检测的 99.8%（SD ± 0.30）和 99.8%（SD ± 0.39）。IH-Int 和 VS-Int 检测的试剂成本/样本分别为 32 美元和 147 美元。所有测序样本均确认为 HIV-1 型 C 群。
IH-Int 检测具有较高的扩增成功率，与商业检测具有高度一致性。与商业检测相比，它的成本显著降低。我们的检测方法具有在博茨瓦纳对基于多替拉韦的方案的参与者进行常规监测所需的规格。
