Morbioli Giorgio Gianini, Baillargeon Keith R, Kalimashe Monalisa N, Kana Vibha, Zwane Hloniphile, van der Walt Cheri, Tierney Allison J, Mora Andrea C, Goosen Mark, Jagaroo Rivashni, Brooks Jessica C, Cutler Ewaldé, Hunt Gillian, Jordan Michael R, Tang Alice, Mace Charles R
Department of Chemistry, Laboratory for Living Devices, Tufts University, Medford, MA 02155.
Center for HIV & Sexually Transmitted Infections, National Institute for Communicable Diseases, Johannesburg 2192, South Africa.
Proc Natl Acad Sci U S A. 2025 Feb 18;122(7):e2419160122. doi: 10.1073/pnas.2419160122. Epub 2025 Feb 10.
Quantifying viral load, a key indicator required to achieve control and elimination of the HIV epidemic, requires cell-free plasma or serum to ensure measurements are not biased by proviral DNA contained in infected CD4 T lymphocytes. Plasma separation cards (PSC) collect and preserve a dried specimen, which makes them practical solutions for decentralized sample collection and transport in limited-resource settings. However, physiological variations in hematocrit levels can introduce significant variability in the quality of plasma generated by commercial PSCs and can lead to inaccurate test results and clinical decisions. In addition to hematocrit-dependent sampling, the Roche PSC, a standard for dried plasma collection, is known to induce considerable hemolysis, which further impacts specimen quality, concordance with liquid plasma, and the overall benefit of microsampling. We address these gaps with a patterned dried plasma spot (pDPS) card, which generates plasma with improved hematocrit independence and minimal hemolysis. This study directly compares pDPS cards to the Roche PSC to measure HIV viral load. Analysis of viral load from 75 donors revealed strong agreement in sensitivity, specificity, overall accuracy, and viral load band placement between devices, with quantitative metrics suggesting improved performance for pDPS cards. In reflexive genotyping, remnant dried blood from pDPS cards exhibited greater success than Roche PSC in amplification and sequencing (71% vs. 62%) and detecting drug resistance mutations (63% vs. 42%). Based on this performance, pDPS cards can be versatile across multiple analytical platforms, integrate seamlessly into existing clinical laboratory workflows, and aid clinicians in making accurate treatment decisions.
量化病毒载量是实现艾滋病疫情控制和消除所需的关键指标，这需要无细胞血浆或血清，以确保测量结果不受感染的CD4 T淋巴细胞中前病毒DNA的影响。血浆分离卡（PSC）可收集并保存干燥样本，这使其成为资源有限环境下分散样本采集和运输的实用解决方案。然而，血细胞比容水平的生理变化会导致商用PSC产生的血浆质量出现显著差异，并可能导致检测结果不准确和临床决策失误。除了依赖血细胞比容的采样外，罗氏PSC作为干燥血浆采集的标准，已知会引起相当程度的溶血，这进一步影响了样本质量、与液态血浆的一致性以及微量采样的整体效益。我们使用图案化干燥血浆点（pDPS）卡来解决这些问题，该卡产生的血浆对血细胞比容的依赖性更小，溶血程度更低。本研究直接将pDPS卡与罗氏PSC进行比较，以测量HIV病毒载量。对75名捐赠者的病毒载量分析显示，两种设备在灵敏度、特异性、总体准确性和病毒载量条带位置方面具有高度一致性，定量指标表明pDPS卡的性能有所提高。在反射基因分型中，pDPS卡残留的干血在扩增和测序（71%对62%）以及检测耐药突变（63%对42%）方面比罗氏PSC更成功。基于这一性能，pDPS卡可在多个分析平台上通用，无缝集成到现有的临床实验室工作流程中，并帮助临床医生做出准确的治疗决策。
