A Microfluidic Device for Label-free HIV Virion Count at Point of Care Settings

用于护理点无标记 HIV 病毒粒子计数的微流体装置

基本信息

批准号：
7860292
负责人：
Xuanhong Cheng
金额：
$ 22.49万
依托单位：
LEHIGH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-05 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7860292
关键词：
Address Adhesions Affect Aluminum Oxide Antibodies Area Back Be++ element Beryllium Binding Biological Clinical Detection Developing Countries Devices Disease Disease Progression Elements Evaluation Event Film Filtration Goals HIV HIV Infections Housing Immobilization Individual Label Life Liquid substance Manuals Measurement Measures Membrane Methods Microfluidic Microchips Microfluidics Modification Molecular Monitor Nucleic Acid Amplification Techniques Nucleic Acids Output Patients Performance Plasma Population Preparation Process Property Protocols documentation Public Health Research Resources Retrieval Sampling Surface System Testing Time Viral Viral Load result Virion Virus base clinical Diagnosis clinically relevant design fluid flow nanoparticle operation particle particle counter point of care pressure public health priorities public health relevance sensor viral detection

项目摘要

DESCRIPTION (provided by applicant): HIV infection affects more than 33 million people globally. Measurement of the level of circulating virus in the blood plasma, or "viral load," is a critical parameter used to monitor disease progression and make treatment decisions. However, it is widely recognized that the standard viral detection approaches, which are based on nucleic acid amplification techniques such as PCR, are technically too demanding and too costly to use at the point of care or in resource-limited settings. A portable, inexpensive and easy-to-operate HIV viral counter is a clinical and public health priority worldwide. To address this need, we propose to develop a microfluidic system for label-free HIV whole particle enumeration from plasma, which avoids the problems associated with sample preparation and nucleic acid amplification, and is suitable for the point of care (POC). To accomplish our goal of measuring circulating virions directly from plasma, we will rely on functional nanoporous anodic aluminum oxide (AAO) membranes packaged in a microfluidic format to filter, concentrate, capture and detect whole HIV virions directly from plasma. We will pursue two specific aims. The first aim intends to develop three microfluidic modules to process virion-containing plasma. The first element is a Virion Separation and Purification Module that will separate HIV virions from other larger particles in the original plasma sample. It will allow >90% viral passage from a 1mL plasma sample in less than 1 minute. The next element is a Viral Concentration Module that will separate smaller particles and enrich the target HIV particles by ~1,000 fold. This concentration will be achieved by packaging appropriately sized, porous AAO membranes in microfluidic devices, to strain plasma and retain HIV virions in the microfluidic chamber above the membrane with minimal viral adhesion. The Viral Concentration Module will accept 1 mL of pre-purified virion-containing plasma as input, and deliver as output <1 <L of an HIV virion-enriched sample suspended in the unpassaged supernatant fraction in less than 4 minutes. The third element is the microfluidic Virion Capture Module. This module will take the concentrated HIV-containing sample as input and immobilize >90% of the target HIV particles on an antibody-functionalized AAO membrane in < 20 minutes, with minimal non-specific binding. In Aim 2, a pressure sensor will be interfaced with the Viral Capture Module across the nanoporous membrane to detect the specific binding events. The pressure build-up resulting from viral immobilization and pore blockage will be measured to calculate viral load in the original sample. In Aim 2, we also plan to integrate all the viral processing components from Aim 1 into a single microfluidic chip and test the collective performance of these components interfaced with the pressure sensor using virus-containing plasma samples. Off-chip valves and LabView interfaces will be designed to control fluid flow and automate chip operation. PUBLIC HEALTH RELEVANCE: Despite its importance in clinical diagnosis and public health, fast and easy approaches for viral detection at the point-of-need are not yet available. The most pressing need for point-of-care viral detection, in the US and globally, is in HIV disease, as most of the 33 million HIV-infected patients have no access to viral load monitoring, which is recommended 3-4 times a year. The current proposal intends to address the challenges by developing a label-free, HIV whole particle counter that can be used at the point of care and in resource limited settings.

描述（由申请人提供）：艾滋病毒感染影响全球超过 3300 万人。测量血浆中循环病毒的水平或“病毒载量”是用于监测疾病进展和做出治疗决策的关键参数。然而，人们普遍认为，基于 PCR 等核酸扩增技术的标准病毒检测方法在技术上要求过高且成本过高，无法在护理点或资源有限的环境中使用。便携式、廉价且易于操作的 HIV 病毒计数器是全球临床和公共卫生的优先事项。为了满足这一需求，我们建议开发一种微流体系统，用于从血浆中进行无标记的 HIV 全颗粒计数，该系统避免了与样品制备和核酸扩增相关的问题，并且适用于护理点 (POC)。为了实现直接从血浆中测量循环病毒体的目标，我们将依靠以微流体形式包装的功能性纳米孔阳极氧化铝 (AAO) 膜来直接从血浆中过滤、浓缩、捕获和检测整个 HIV 病毒体。我们将追求两个具体目标。第一个目标是开发三种微流体模块来处理含有病毒颗粒的血浆。第一个元件是病毒粒子分离和纯化模块，它将 HIV 病毒粒子与原始血浆样本中的其他较大颗粒分离。它可以在不到 1 分钟的时间内从 1mL 血浆样本中实现 >90% 的病毒通过。下一个元件是病毒浓缩模块，它将分离较小的颗粒并将目标 HIV 颗粒富集约 1,000 倍。这种浓度将通过在微流体装置中包装适当尺寸的多孔 AAO 膜来实现，以过滤血浆并将 HIV 病毒粒子保留在膜上方的微流体室中，同时将病毒粘附降至最低。病毒浓缩模块将接受 1 mL 预纯化的含病毒粒子的血浆作为输入，并在不到 4 分钟内输出悬浮在未传代的上清液部分中的 <1 <L 的富含 HIV 病毒粒子的样品。第三个元件是微流体病毒粒子捕获模块。该模块将以浓缩的含有 HIV 的样品作为输入，并在 20 分钟内将超过 90% 的目标 HIV 颗粒固定在抗体功能化的 AAO 膜上，同时将非特异性结合降至最低。在目标 2 中，压力传感器将通过纳米多孔膜与病毒捕获模块连接，以检测特定的结合事件。将测量由病毒固定和孔堵塞引起的压力积聚，以计算原始样品中的病毒载量。在目标 2 中，我们还计划将目标 1 中的所有病毒处理组件集成到单个微流控芯片中，并使用含病毒的血浆样本测试与压力传感器连接的这些组件的集体性能。片外阀门和 LabView 接口将被设计用于控制流体流动和自动化芯片操作。公共卫生相关性：尽管病毒检测在临床诊断和公共卫生中很重要，但目前还没有快速简便的病毒检测方法。在美国和全球范围内，对即时病毒检测最迫切的需求是 HIV 疾病，因为 3300 万 HIV 感染者中的大多数无法进行病毒载量监测，建议每年进行 3-4 次病毒载量监测。年。目前的提案旨在通过开发一种无标签的艾滋病毒全颗粒计数器来应对这些挑战，该计数器可在护理点和资源有限的环境中使用。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Micropatterned macroporous structures in microfluidic devices for viral separation from whole blood.

微流体装置中的微图案大孔结构，用于从全血中分离病毒。

DOI：
10.1039/c7an00576h
发表时间：
2017
期刊：
The Analyst
影响因子：
0
作者：
Surawathanawises,Krissada;Wiedorn,Victoria;Cheng,Xuanhong
通讯作者：
Cheng,Xuanhong

Microfluidic devices with templated regular macroporous structures for HIV viral capture.

具有模板化规则大孔结构的微流体装置，用于捕获 HIV 病毒。

DOI：
10.1039/c5an02282g
发表时间：
2016
期刊：
The Analyst
影响因子：
0
作者：
Surawathanawises,Krissada;Kundrod,Kathryn;Cheng,Xuanhong
通讯作者：
Cheng,Xuanhong

Measuring the Soret coefficient of nanoparticles in a dilute suspension.

测量稀悬浮液中纳米颗粒的索雷系数。

DOI：
10.1007/s11051-014-2625-6
发表时间：
2014
期刊：
Journal of nanoparticle research : an interdisciplinary forum for nanoscale science and technology
影响因子：
0
作者：
Zhao,Chao;Fu,Jinxin;Oztekin,Alparslan;Cheng,Xuanhong
通讯作者：
Cheng,Xuanhong

Gravity-induced swirl of nanoparticles in microfluidics.