Acoustofluidic Salivary Exosome Isolation A Liquid Biopsy Compatible Approach for Human Papillomavirus-Associated Oropharyngeal Cancer Detection
Acoustofluidic Salivary Exosome Isolation A Liquid Biopsy Compatible Approach for Human Papillomavirus-Associated Oropharyngeal Cancer Detection
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DOI：
10.1016/j.jmoldx.2019.08.004
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发表时间：
2020-01-01
影响因子：
4.1
通讯作者：
Huang, Tony Jun
中科院分区：
文献类型：
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作者：
Wang, Zeyu;Li, Feng;Huang, Tony Jun
以前的努力，以评估检测人乳头瘤病毒（HPV）DNA在整个唾液作为诊断措施HPV相关口咽癌（HPV-OPC）没有显示出足够的临床性能。我们假设唾液外泌体与HPV相关的生物标志物一起包装，并且通过分离有效富集唾液外泌体可以增强HPV-OPC的诊断和预后性能。在这项研究中，开发了一种声流体（声学和微流体的融合）平台来进行基于大小的唾液外泌体分离。这些数据表明，无论粘度变化和收集方法如何，该平台都能够始终如一地从唾液样品中分离外泌体。与目前的金标准相比，差速离心、液滴数字RT-PCR分析表明，来自声流平台的唾液外泌体小RNA的平均产量高出15倍。利用这种高产量的外泌体分离平台，我们表明可以在从HPV相关OPC患者的唾液中分离的外泌体中检测到HPV 16 DNA，与HPV 16阳性的组织/活检的一致性为80%。总的来说，这些数据表明，声流体平台可以实现高纯度和高产量的唾液外泌体分离，用于下游基于唾液外泌体的液体活检应用。此外，HPV-OPC患者中的HPV 16 DNA序列包装在唾液外泌体中，它们的分离将增强HPV 16 DNA的检测。
Previous efforts to evaluate the detection of human papilloma viral (HPV) DNA in whole saliva as a diagnostic measure for H PV-associated oropharyngeal cancer (HPV-OPC) have not shown sufficient clinical performance. We hypothesize that salivary exosomes are packaged with HPV-associated biomarkers, and efficient enrichment of salivary exosomes through isolation can enhance diagnostic and prognostic performance for HPV-OPC. In this study, an acoustofluidic (the fusion of acoustics and microfluidics) platform was developed to perform size-based isolation of salivary exosomes. These data showed that this platform is capable of consistently isolating exosomes from saliva samples, regardless of viscosity variation and collection method. Compared with the current gold standard, differential centrifugation, droplet digital RT-PCR analysis showed that the average yield of salivary exosomal small RNA from the acoustofluidic platform is 15 times higher. With this high-yield exosome isolation platform, we show that H PV16 DNA could be detected in isolated exosomes from the saliva of HPV-associated OPC patients at 80% concordance with tissues/biopsies positive for HPV16. Overall, these data demonstrated that the acoustofluidic platform can achieve high-purity and high-yield salivary exosome isolation for downstream salivary exosome based liquid biopsy applications. Additionally, HPV16 DNA sequences in HPV-OPC patients are packaged in salivary exosomes and their isolation will enhance the detection of HPV16 DNA.