High sensitivity detection and sorting of infectious human immunodeficiency virus (HIV-1) particles by flow virometry
High sensitivity detection and sorting of infectious human immunodeficiency virus (HIV-1) particles by flow virometry
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DOI：
10.1016/j.virol.2017.02.016
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发表时间：
2017-05-01
期刊：
影响因子：
3.7
通讯作者：
Tilton, John C.
中科院分区：
文献类型：
--
作者：
Bonar, Michal M.;Tilton, John C.
通过流式细胞术检测病毒由于其小尺寸而变得复杂。在这里，我们表征了标准（FACSAria II）和亚微米流式细胞仪（A50 Micro）分辨HIV-1病毒的能力。A50在分辨小颗粒方面具有上级优势，但仅通过光散射特性不能可靠地区分HIV-1、细胞外囊泡和激光噪声。然而，单荧光HIV-1颗粒可以很容易地检测到两个细胞仪。荧光颗粒被分选并保留感染性，允许进一步探索HIV-1异质性的功能后果。最后，流式细胞术的检测限为80个病毒/ml，几乎等于PCR检测。这些研究证明了流式细胞术检测和分选病毒颗粒的能力，并提供了一个关键的工具包来验证标记野生型HIV-1的方法;定量评估病毒和基于病毒的治疗剂的完整性和聚集性;并有效筛选抑制病毒组装和释放的药物。
Detection of viruses by flow cytometry is complicated by their small size. Here, we characterized the ability of a standard (FACSAria II) and a sub-micron flow cytometer (A50 Micro) to resolve HIV-1 viruses. The A50 was superior at resolving small particles but did not reliably distinguish HIV-1, extracellular vesicles, and laser noise by light scatter properties alone. However, single fluorescent HIV-1 particles could readily be detected by both cytometers. Fluorescent particles were sorted and retained infectivity, permitting further exploration of the functional consequences of HIV-1 heterogeneity. Finally, flow cytometry had a limit of detection of 80 viruses/ml, nearly equal to PCR assays. These studies demonstrate the power of flow cytometry to detect and sort viral particles and provide a critical toolkit to validate methods to label wild-type HIV-1; quantitatively assess integrity and aggregation of viruses and virus-based therapeutics; and efficiently screen drugs inhibiting viral assembly and release.