Novel Viral Load Diagnostic for Resource-Limited Setting

描述（由申请人提供）：缺乏负担得起的和实用的艾滋病毒诊断，包括监测艾滋病毒RNA水平（“病毒载量”）和诊断婴儿艾滋病毒感染的测定，已被确定为全球艾滋病毒护理的主要障碍。对于资源有限的环境，一个有用的HIV RNA诊断检测将是简单的，自动化的，快速的，每次测试的成本低于10美元。我们建议使用BioScale的专有传感器格式开发符合这些规格的HIV病毒体检测。 我们假设，用特异性生理受体（DC-SIGN，CD 4）和/或针对表面表位的新单克隆抗体捕获整个病毒体，并将其整合到BioScale检测平台上的检测中，将提供一种灵敏的新诊断方法，满足资源匮乏环境的要求。在该项目的第一年，捕获表面将使用重组可溶性CD 4，重组可溶性DC-SIGN和新的抗gp 41和抗gp 120抗体进行功能化，以创建一种新的系统来捕获和分离HIV病毒体并将其锚在传感器表面。在第二年，这种新的HIV病毒体检测将在75名HIV感染者中进行优化，并与标准病毒载量测量（即RT-PCR）相比，验证其灵敏度和特异性。该提案的成功完成将证明BioScale的HIV检测在资源有限的环境中的可行性。 该提案的具体目标有三个方面： （A）开发一种使用包被有受体和/或新的单克隆抗体的磁珠捕获HIV病毒体的方法; B）在BioScale生物传感器上开发珠捕获表面并将其与HIV捕获珠整合，以创建HIV病毒体测定;以及 C）在HIV感染受试者中，评价通过HIV测定检测与RT-PCR相比在全血中HIV检测的定性和定量阈值。 该提案的第二阶段预计将包括仪器的β测试、更大规模的临床测试以及仪器和检测试剂盒制造的准备和在资源有限环境中的部署。

DESCRIPTION (provided by applicant): The lack of affordable and practical HIV diagnostics, including assays to monitor HIV RNA levels ("viral load") and to diagnose HIV infection in infants, has been identified as a major barrier to HIV care worldwide. For resource-limited settings, a useful HIV RNA diagnostic assay would be simple, automated, and rapid and cost less than $10 per test. We propose to develop an HIV virion assay that meets these specifications using BioScale's proprietary sensor format. We hypothesize that capturing whole virions with specific physiological receptors (DC-SIGN, CD4) and/or new monoclonal antibodies against surface epitopes and integrating them into an assay on BioScale's detection platform will provide a sensitive new diagnostic that meets requirements of resource-poor setting. In the first year of the project, capture surfaces will be functionalized using recombinant soluble CD4, recombinant soluble DC-SIGN, and new anti-gp41 and anti-gp120 antibodies to create a novel system to capture and isolate HIV virions and anchor them to the sensor surface. In the second year, this new HIV virion assay will be optimized and validated for sensitivity and specificity compared to standard viral load measurements (i.e. RT-PCR) in 75 HIV-infected subjects. The successful completion of this proposal will demonstrate the feasibility of BioScale's HIV assay for resource-limited settings. The specific aims of the proposal are three-fold: A) To develop a method to capture HIV virions using magnetic beads coated with receptors and/or new monoclonal antibodies; B) To develop a bead capture surface on the BioScale biosensor and integrate it with HIV capture beads, to create an HIV virion assay; and C) To evaluate the qualitative and quantitative threshold of HIV detection in whole blood via HIV assay testing compared to RT-PCR in HIV-infected subjects. The Phase II of this proposal is anticipated to include beta-testing of the instrument, larger clinical testing, and preparation for instrument and assay kit manufacturing and deployment in resource-limited settings.