Nanopore-Based HIV Self-Test for Ultrasensitive p24 Quantification in FingerPrick Blood

基于纳米孔的 HIV 自检，可对 FingerPrick 血液中的 p24 进行超灵敏定量

基本信息

批准号：
10594132
负责人：
Chang Liu
金额：
$ 41.3万
依托单位：
UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-18 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10594132
关键词：
AIDS prevention Acute Adherence Age Antibody Response Antigens Belief Benchmarking Benefits and Risks Biological Assay Biological Markers Blood Blood Glucose Blood specimen CD4 Lymphocyte Count Cellular Phone Characteristics Chemistry Clinical Clinical Markers Clinical assessments Detection Devices Diagnostic Drug resistance Early Diagnosis Early identification Enrollment Ensure Enzyme-Linked Immunosorbent Assay Feedback Fingers Generations HIV HIV Antigens HIV Infections HIV Seronegativity HIV-1 HIV/AIDS Health Home Human immunodeficiency virus test Immunology Incidence Individual Infection Ions Label Laboratories Learning Longitudinal Studies Measurement Measures Mediating Methods Microfluidic Microchips Microfluidics Monitor Nucleic Acid Amplification Tests Nucleic Acids Participant Patients Pattern Performance Persons Phase Polymerase Chain Reaction Population Preparation Privatization Process Proteins Proteomics Protocols documentation RNA Reaction Reader Research Risk Sampling Sensitivity and Specificity Signal Transduction Specificity Specimen Structure Surveys Technology Testing Therapeutic Time Treatment Efficacy Treatment Failure United States Validation Viral Load result Whole Blood acute infection antibody detection antigen challenge antigen detection antigen test antiretroviral therapy cost cost effective detection limit detection sensitivity experience improved innovation isothermal amplification minimally invasive nanoparticle nanopore nanosized performance tests prisma prototype recruit research and development research clinical testing satisfaction self testing sex therapy adherence transmission process usability user friendly software viral detection viral rebound

项目摘要

ABSTRACT Despite tremendous advances in treatment of HIV/AIDS and the decrease of HIV incidence, the overall infected population continues to grow. Progress on prevention of HIV transmission remains far too slow. It is estimated that 20% of new HIV infections are due to transmission from unaware infected individuals. Hence, early detection of HIV is particularly important for lowering transmission rates. To this end, extending testing accessibility beyond clinical settings through self-tests is highly desirable. HIV self-testing is a process in which an individual who wants to know his/her HIV status collects a specimen, performs the test, and interprets the result in private. Current HIV self-testing technologies include rapid protein tests and nucleic acid tests. The suboptimal sensitivity of current protein tests can only support antibody detection and will miss a significant portion of acute infections. Although nucleic acid tests can reach lower detection limit through amplification technologies such as PCR, they will miss the information that antigens can provide. Most current research efforts on HIV self-testing are focused on nucleic acid tests. However, there is no evidence showing that HIV RNA appears ahead of antigen. The major technological challenge for antigen detection is that proteins cannot be amplified like nucleic acids, leading to the widely held belief that antigen tests are relatively insensitive and therefore have a limited clinical utility. We previously demonstrated a click chemistry amplified nanopore (CAN) assay method for ultrasensitive antigen quantification. This assay achieved 0.5 pg/ml detection limit for HIV-1 p24 antigen and demonstrated reliable detection in clinical samples from patients missed by nucleic acid and/or ELISA assays. Quantitative p24 results from this method also indicated correlation between p24 and viral load, suggesting potential use for monitoring antiretroviral therapy adherence to minimize treatment failure. Based on the CAN assay, this project aims to develop an ultrasensitive quantitative HIV-1 p24 antigen self-test to improve early detection of acute infections and monitoring treatment efficacy. The test will be based on a streamlined automatic device including a cost- effective microfluidic chip for sample preparation and a nanopore reader for laypersons to test themselves using fingerprick blood. The R61 phase will develop a self-testing device for quantification of HIV-1 p24 antigen at ≤ 2 pg/ml in whole blood to support detection of acute infection and treatment failure. The R33 phase will perform a clinical evaluation at the Prisma Health Immunology Center to assess the performance, usability, and acceptability of the self-testing device. Through innovations in click chemistry amplified nanopore detection and microfluidic sample preparation, we anticipate the test would be able to quantify HIV-1 p24 antigen at as low as 0.5 pg/ml directly from 100µl or less finger prick blood and establish correlation between p24 and viral load levels. If successfully developed and validated, this CAN self-testing device should enable routine HIV self-testing as simple as a blood sugar test at home to support global HIV diagnostic and therapeutic efforts.

抽象的尽管艾滋病毒/艾滋病的治疗取得了巨大进步，艾滋病毒感染率有所下降，但总体感染人数据估计，预防艾滋病毒传播的进展仍然十分缓慢。 20％的新艾滋病毒感染是由于不知情的感染者传播的，因此，早期发现。为此，扩大检测范围对于降低传播率尤为重要。临床环境中通过自我检测是非常可取的。 HIV 自我检测是一个个人进行自我检测的过程。如果想了解自己的艾滋病毒状况，可以私下采集样本、进行检测并解读结果。目前的艾滋病毒自我检测技术包括快速蛋白质检测和核酸检测，灵敏度欠佳。目前的蛋白质检测只能支持抗体检测，并且会漏掉很大一部分急性感染。虽然核酸检测可以通过PCR等扩增技术达到较低的检出限，但将会错过抗原可以提供的信息。目前大多数关于艾滋病毒自我检测的研究工作都集中在这一点上。然而，没有证据表明HIV RNA出现在抗原之前。抗原检测的技术挑战是蛋白质不能像核酸一样被扩增，从而导致人们普遍认为抗原测试相对不敏感，因此临床用途有限。我们之前演示了用于超灵敏抗原的点击化学放大纳米孔 (CAN) 测定方法该检测方法实现了 HIV-1 p24 抗原的 0.5 pg/ml 检测限，并证明是可靠的。核酸和/或 ELISA 定量检测漏掉的患者临床样本中的检测。该方法还表明 p24 和病毒载量之间的相关性，表明其可能用于监测该项目旨在提高抗逆转录病毒治疗的依从性，以最大程度地减少治疗失败。开发超灵敏定量 HIV-1 p24 抗原自检，以改善急性感染的早期检测该测试将基于一个简化的自动设备，包括成本- 用于样品制备的有效微流控芯片和供外行人员自行测试的纳米孔读取器 R61阶段将开发一种自检装置，用于定量≤2的HIV-1 p24抗原。全血中的 pg/ml，以支持急性感染和治疗失败的检测。 Prisma Health 免疫学中心进行临床评估，以评估性能、可用性和通过点击化学放大纳米孔检测和技术的创新，自检设备的可接受性。微流体样品制备，我们预计该测试能够以低至直接从 100 µl 或更少的手指刺血中提取 0.5 pg/ml，并建立 p24 和病毒载量水平之间的相关性。如果成功开发和验证，这种 CAN 自检设备应该能够进行常规 HIV 自检就像在家进行血糖测试一样简单，以支持全球艾滋病毒诊断和治疗工作。