Quantitative microfluidic NAT-on-USB: towards routine HIV viral load testing

定量微流控 NAT-on-USB：走向常规 HIV 病毒载量检测

• 批准号: 1912410
• 负责人: Weihua Guan
• 金额: $ 35.25万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1912410&HistoricalAwards=false
• 关键词: Quantitative; microfluidic; NAT; USB; towards

HIV, the virus that causes AIDS, continues to be a significant public health issue. Fortunately, HIV can be effectively managed if people of HIV can be identified and their virus is controlled to undetectable levels by antiretroviral therapy. Increasing access to routine viral load testing can help reduce the treatment failure rate by early detection of viral rebound caused by either drug resistance or poor therapy adherence. To this end, HIV self-testing, a process in which individual who wants to know HIV status collects a specimen, performs a test and interprets the result in private, has become an empowering and innovative approach. Existing HIV self-testing methods rely almost exclusively on lateral flow based test to detect host antibody response to HIV infection. They could miss a significant portion of asymptomatic individuals during the 6-12 weeks of early infection window; they also lack the ability for detection of viral rebound. Nucleic acid testing is currently the only method for viral load quantification. Nevertheless, its use by laypersons is limited due to the sample handling and assay complexity. The proposed research has the potential to significantly enhance the treatment outcomes for individuals of HIV under antiretroviral therapy. In addition, with educational and outreach activities, the proposed research is well positioned to enhance the integrative learning experience and to engage multidisciplinary students at many levels.The primary research objective of this proposal is to explore an ultra-compact quantitative nucleic acid testing (NAT) on a disposable microfluidic chip with a USB analyzer to detect HIV-1 viral rebound that is simple enough for laypersons to test themselves to monitor treatment adherence. The goal is to develop and validate a whole blood-based test that can semi-quantitatively assess the presence of HIV-1 RNA at cell concentrations as low as 1000 copies/ml. The test can be performed similarly to a home blood glucose test with a single step of finger-prick blood loading. To this end, four research tasks regarding the microfluidic sample preparation, amplification assay, USB analyzer integration, and analytical validation will be pursued. Task 1. Automated microfluidic sample preparation. A streamlined microfluidic chip for automated plasma separation and RNA extraction from whole blood will be developed. Task 2. An HIV-1 Real-time reverse transcription loop-mediated isothermal amplification assay will be optimized and the lowest possible limit of detection will be explored. Task 3. Analyzer integration and small-scale prototyping. The analyzer hardware and software will be developed and integrated for easy and robust operation. Task 4. Laboratory validation with control samples. The prototyped device will be validated in lab settings using HIV-1 plasma samples spiked into whole blood.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

引起艾滋病的病毒艾滋病毒仍然是一个重大的公共卫生问题。幸运的是，如果可以鉴定出艾滋病毒的人并通过抗逆转录病毒疗法将艾滋病毒的病毒控制为无法检测到的水平，则可以有效地管理艾滋病毒。增加对常规病毒负荷测试的机会可以通过早期发现由耐药性或治疗不良引起的病毒反弹来帮助降低治疗失败率。为此，艾滋病毒自我测试是一个想知道艾滋病毒状况的个人收集标本，执行测试并将结果解释为私人的过程，已成为一种授权和创新的方法。现有的HIV自我测试方法几乎完全依赖于基于横向流的测试来检测宿主抗体对HIV感染的反应。在早期感染窗口的6-12周内，他们可能会错过大部分无症状的个体；他们还缺乏检测病毒反弹的能力。核酸测试目前是病毒负荷定量的唯一方法。然而，由于样品处理和测定复杂性，外行人的使用受到限制。拟议的研究有可能在抗逆转录病毒疗法下显着增强HIV患者的治疗结果。此外，借助教育和宣传活动，拟议的研究非常适合增强综合学习经验并在许多级别上吸引多学科的学生。该提案的主要研究目标是探索超紧凑型核酸测试（NAT）在一次性的usb分析者中，以对HIV的分析仪进行较简单的研究，以使HIV的杂物对HIV的反驳，以使HIV探测HIV的杂物。目的是开发和验证全血测试，该测试可以在细胞浓度低至1000拷贝/mL的细胞浓度下半量化评估HIV-1 RNA的存在。该测试可以类似于家庭血糖测试，并具有单一的手指刺激性。为此，将采用有关微流体样本制备，放大测定，USB分析仪集成和分析验证的四项研究任务。任务1。自动微流体样品制备。将开发出简化的微流体芯片，用于自动血浆分离和从全血中提取RNA。任务2。将优化HIV-1实时逆转录环介导的等温放大测定法，并将探索最低的检测限制。任务3。分析仪集成和小规模的原型制作。分析仪硬件和软件将被开发和集成，以便于轻松，强大的操作。任务4。实验室验证使用控制样品。该原型设备将在实验室设置中使用HIV-1等离子体样本进行验证。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的智力优点和更广泛的影响评估评估标准通过评估来支持的。

项目成果

Nanopore Digital Counting of Amplicons for Ultrasensitive Electronic DNA Detection

用于超灵敏电子 DNA 检测的扩增子纳米孔数字计数

• DOI: 10.1109/iedm19573.2019.8993671
• 发表时间: 2019
• 期刊: 2019 IEEE International Electron Devices Meeting (IEDM
• 作者: Tang, Zifan;Choi, Gihoon;Nouri, Reza;Guan, Weihua
• 通讯作者: Guan, Weihua

Digital CRISPR Systems for the Next Generation of Nucleic Acid Quantification.

• DOI: 10.1016/j.trac.2023.116917
• 发表时间: 2023-01
• 期刊: Trends in analytical chemistry : TRAC
• 作者: Anthony J Politza;Reza Nouri;W. Guan

Fingerpick Blood-Based Nucleic Acid Testing on A USB Interfaced Device towards HIV self-testing.

• DOI: 10.1016/j.bios.2022.114255
• 发表时间: 2022-08-01
• 期刊: Biosensors & bioelectronics
• 影响因子: 12.6

Sensitive and specific CRISPR-Cas12a assisted nanopore with RPA for Monkeypox detection.

• DOI: 10.1016/j.bios.2023.115866
• 发表时间: 2023-11
• 期刊: Biosensors & bioelectronics
• 影响因子: 12.6
• 作者: Md. Ahasan Ahamed;Muhammad Asad Ullah Khalid;Ming Dong;Anthony J Politza;Zhikun Zhang;Aneesh Kshirsagar;Tianyi Liu;W. Guan

Programmable Magnetic Robot (ProMagBot) For Fully Automated Nucleic Acid Sample Preparation at The Point of Need

可编程磁性机器人 (ProMagBot) 用于在需要时全自动制备核酸样品

• 发表时间: 2022
• 期刊: Hilton Head Workshop
• 作者: A. J. Politza, T. Liu