BACKGROUND 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. METHODS 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. FINDINGS 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. INTERPRETATION 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.

中文翻译：

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OLA-Simple：针对低资源实验室的软件指导的HIV-1耐药性测试。

背景技术HIV耐药性（HIVDR）测试可以帮助临床医生选择治疗方法。然而，基因分型分析的高复杂性和成本限制了在HIVDR患病率已达到很高水平的环境中进行的常规检测。方法开发了寡核苷酸连接测定法（OLA）-简单试剂盒，用于检测针对一线非核苷/核苷逆转录酶抑制剂的HIVDR，并在来自A，B，C，D和AE亚型的672个密码子（168个标本）上进行了验证。该套件使用干燥试剂来促进测定设置，用于视觉HIVDR检测的侧向流动装置以及带有用于指导用户和分析结果的界面的内部软件。结果与Sanger测序相比，通过OLA-Simple对标本进行的HIVDR分析显示出99.6±0.3％的特异性和98.2±0.9％的灵敏度，而与高灵敏度的测定相比则为99。特异性为6±0.6％，灵敏度为86.2±2.5％，不确定结果为2.6±0.9％。与Sanger测序相比，OLA-Simple的执行速度更快（<4小时vs. 35-72小时）。41名未经训练的志愿者盲目测试了两个标本，每个标本的准确度为96.8±0.8％。解释OLA-Simple与Sanger和敏感的比较者进行的HIVDR基因分型相比具有优势。教学软件使没有经验的新手可以高精度地执行测定。OLA-Simple的降低的复杂性，成本和培训要求可以改善在资源贫乏地区获得HIVDR测试的机会，并有可能允许在当天选择适当的抗逆转录病毒疗法。资金：美国国立卫生研究院R01；UW CFAR的临床和逆转录病毒学研究核心以及分子谱和计算生物学核心；西雅图儿童研究所；UW Holloman创新挑战奖；皮尔彻学院奖学金。