Designing and Interpreting Limiting Dilution Assays: General Principles and Applications to the Latent Reservoir for Human Immunodeficiency Virus-1.
Designing and Interpreting Limiting Dilution Assays: General Principles and Applications to the Latent Reservoir for Human Immunodeficiency Virus-1.
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DOI：
10.1093/ofid/ofv123
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发表时间：
2015-12
影响因子：
4.2
通讯作者：
Siliciano RF
中科院分区：
文献类型：
--
作者：
Rosenbloom DI;Elliott O;Hill AL;Henrich TJ;Siliciano JM;Siliciano RF
有限稀释法广泛应用于传染病研究。这些检测方法对目前的HIV-1治疗研究至关重要。在这里，我们提供了新的工具，以帮助研究人员根据他们的特定研究需求设计和分析稀释测定。有限稀释法广泛应用于传染病研究。这些检测方法对于当前人类免疫缺陷病毒（HIV）-1治疗研究至关重要。在这项研究中，我们提供了新的工具，以帮助研究人员设计和分析稀释测定基于他们的特定研究需求。有限稀释试验通常用于测量感染程度，在HIV背景下，它们代表了研究潜伏期和潜在治疗策略的重要工具。然而，标准的检测设计可能无法辨别干预是否会减少已经很小的潜伏感染。这篇评论解决了在这种情况下，并在一般使用的稀释测定中出现的挑战。我们说明了主要的统计方法估计频率的传染性单位从化验结果，我们提供了一个在线工具来计算这个估计。我们建议一个程序，定制检测设计，以实现所需的灵敏度和精度目标，受实验约束。我们认为实验中没有观察到病毒的生长，并解释如何使用替代品的病毒生长可能使测量HIV潜伏期更有效。最后，我们讨论了生物并发症，如小感染的概率增长，改变实验结果的解释。
Limiting dilution assays are widely used in infectious disease research. These assays are crucial for current HIV-1 cure research in particular. Here we offer new tools to help investigators design and analyze dilution assays based on their specific research needs. Limiting dilution assays are widely used in infectious disease research. These assays are crucial for current human immunodeficiency virus (HIV)-1 cure research in particular. In this study, we offer new tools to help investigators design and analyze dilution assays based on their specific research needs. Limiting dilution assays are commonly used to measure the extent of infection, and in the context of HIV they represent an essential tool for studying latency and potential curative strategies. Yet standard assay designs may not discern whether an intervention reduces an already miniscule latent infection. This review addresses challenges arising in this setting and in the general use of dilution assays. We illustrate the major statistical method for estimating frequency of infectious units from assay results, and we offer an online tool for computing this estimate. We recommend a procedure for customizing assay design to achieve desired sensitivity and precision goals, subject to experimental constraints. We consider experiments in which no viral outgrowth is observed and explain how using alternatives to viral outgrowth may make measurement of HIV latency more efficient. Finally, we discuss how biological complications, such as probabilistic growth of small infections, alter interpretations of experimental results.