Department of Bioengineering, University of Washington, Seattle, WA 98195, USA; Global WACh Program, Department of Global Health, University of Washington, Seattle, WA 98104, USA.
Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, WA 98109, USA.
EBioMedicine. 2019 Dec;50:34-44. doi: 10.1016/j.ebiom.2019.11.002. Epub 2019 Nov 22.
HIV drug resistance (HIVDR) testing can assist clinicians in selecting treatments. However, high complexity and cost of genotyping assays limit routine testing in settings where HIVDR prevalence has reached high levels.
The oligonucleotide ligation assay (OLA)-Simple kit was developed for detection of HIVDR against first-line non-nucleoside/nucleoside reverse transcriptase inhibitors and validated on 672 codons (168 specimens) from subtypes A, B, C, D, and AE. The kit uses dry reagents to facilitate assay setup, lateral flow devices for visual HIVDR detections, and in-house software with an interface for guiding users and analyzing results.
HIVDR analysis of specimens by OLA-Simple compared to Sanger sequencing revealed 99.6 ± 0.3% specificity and 98.2 ± 0.9% sensitivity, and compared to high-sensitivity assays, 99.6 ± 0.6% specificity and 86.2 ± 2.5% sensitivity, with 2.6 ± 0.9% indeterminate results. OLA-Simple was performed more rapidly compared to Sanger sequencing (<4 h vs. 35-72 h). Forty-one untrained volunteers blindly tested two specimens each with 96.8 ± 0.8% accuracy.
OLA-Simple compares favorably with HIVDR genotyping by Sanger and sensitive comparators. Instructional software enabled inexperienced, first-time users to perform the assay with high accuracy. The reduced complexity, cost, and training requirements of OLA-Simple could improve access to HIVDR testing in low-resource settings and potentially allow same-day selection of appropriate antiretroviral therapy. FUND: USA National Institutes of Health R01; the Clinical and Retrovirology Research Core and the Molecular Profiling and Computational Biology Core of the UW CFAR; Seattle Children's Research Institute; UW Holloman Innovation Challenge Award; Pilcher Faculty Fellowship.
HIV 耐药性（HIVDR）检测可以帮助临床医生选择治疗方案。然而，基因分型检测的复杂性和成本较高，限制了在 HIVDR 流行率已经达到较高水平的环境中进行常规检测。
开发了寡核苷酸连接检测（OLA）-Simple 试剂盒，用于检测针对一线非核苷/核苷逆转录酶抑制剂的 HIVDR，并在 A、B、C、D 和 AE 亚型的 672 个密码子（168 个标本）上进行了验证。该试剂盒使用干试剂来方便检测设置，使用侧向流动设备进行直观的 HIVDR 检测，并使用内部软件，具有指导用户和分析结果的界面。
OLA-Simple 与 Sanger 测序相比，对标本的 HIVDR 分析显示出 99.6±0.3%的特异性和 98.2±0.9%的敏感性，与高灵敏度检测相比，特异性为 99.6±0.6%，敏感性为 86.2±2.5%，不确定结果为 2.6±0.9%。OLA-Simple 比 Sanger 测序更快完成（<4 小时 vs. 35-72 小时）。41 名未经训练的志愿者对每个样本进行了两次盲测，准确率为 96.8±0.8%。
OLA-Simple 与 Sanger 基因分型和敏感比较器相比具有优势。有指导的软件使无经验的首次使用者能够以高准确度进行检测。OLA-Simple 的简化复杂性、成本和培训要求可以提高在资源匮乏环境中获得 HIVDR 检测的机会，并有可能允许当天选择适当的抗逆转录病毒治疗。资金来源：美国国立卫生研究院 R01；UW CFAR 的临床和逆转录病毒研究核心以及分子分析和计算生物学核心；西雅图儿童研究所；UW 霍洛曼创新挑战赛奖；皮勒尔教席奖学金。
