Rapid point of care HIV PCR diagnostic device

描述(由申请人提供)：自20世纪80年代末以来，确认HIV-1感染的国家指南规定使用免疫印迹(WB)和间接免疫荧光分析(IFA)进行诊断1。然而，核酸扩增测试(NAAT)可能会更早地发现一些感染；降低测试成本；允许进行HIV-2和更广泛的HIV-1亚型检测；减少提供假阳性、假阴性和不确定的测试结果。目前不建议使用NAAT进行筛查，因为它的成本要高得多，而且涉及复杂的方法学。然而，在护理地点进行低成本、快速的NAAT检测在性传播感染(STI)诊所或急诊室(患者通常不会回来查看检测结果)或之前未接受检测的高危孕妇的产房和分娩病房中将是有用的。在所有卫生保健环境中进行更频繁的筛查可能会增加获知结果并可与适当护理联系起来的艾滋病毒感染者的比例。Microronic的目标是开发一种用于艾滋病毒前病毒DNA检测的首个同类、低成本的定性PCR检测方法，该方法在封闭的系统、一次性设备内实现自动化，并允许在靠近患者护理地点和低资源环境下处理样本输入/结果输出。其意图是以低于5美元的价格出售这款设备，而目前NAAT测试的成本为45至75美元。检测策略如下：使用层压聚合物装置内的流体和阀逻辑从全血指棒(20-50微升)样本中提取DNA，并将洗脱出的核酸直接引入微通道以对干燥的PCR母体混合物进行再水合。在不到10分钟的时间内，在微通道内快速进行PCR。扩增产物用FRET方法进行荧光标记，并在40个热循环结束时光学检测到阳性结果。在设备提取和扩增之后，成功完成第一阶段10个拷贝HIV检测的里程碑将证实我们的检测集成和关闭设备策略的有效性。 与公共卫生相关：为了更好地识别和咨询未被确认的艾滋病毒感染者，并将他们与临床和预防服务联系起来，以及进一步减少美国艾滋病毒的围产期传播，疾控中心建议扩大筛查。最灵敏的筛查方法是基于对核酸检测的检测，但这种方法目前成本很高，而且是在参考实验室进行的。我们提出了核酸检测方法的小型化和自动化，以实现低成本、接近患者的检测。

DESCRIPTION (provided by applicant): Since the late 1980s national guidelines to confirm HIV-1 infection dictate the use of Western blot (WB) and indirect immunofluorescence assay (IFA) for diagnosis1. However, nucleic acid amplification tests (NAAT) may identify some infections earlier; reduce testing costs; permit HIV-2 and broader HIV-1 subtype detection; reduce provision of false-positive, false-negative, and indeterminate test results. NAAT is currently not recommended for screening since it is much more expensive and involves complex methodology. However, a low cost, rapid NAAT test at point of care would be useful in Sexually Transmitted Infection (STI) clinics or emergency rooms (where patients often do not return for tests results) or on labor and delivery wards for high-risk pregnant women who have not previously been tested. More frequent screening in all health care settings could increase the proportion of HIV-infected clients who learn their results and who could be linked to appropriate care. Micronic's goal is to develop a firstof- its-kind, low cost qualitative PCR assay for HIV provirus DNA detection that is automated within a closed system, disposable device and allows specimen in / result out processing at near patient point of care and in low resource settings. The intent is to sell this device at under $5 as compared to the $45 to $75 current NAAT tests cost. The assay strategy is as follows : DNA is extracted from a whole blood finger stick (20-50 microliter) specimen using fluidic and valve logic within a laminate polymer device and the eluted nucleic acid introduced directly into microchannels to rehydrate a dried PCR master mix. PCR is rapidly performed within the microchannels in under 10 minutes. The amplified product is fluorescently tagged by a FRET method and a positive result detected optically at the end of 40 thermal cycles. Successful completion of the phase I milestone of 10 copy HIV detection following device extraction and amplification will confirm the validity of our assay integration and closed device strategy. PUBLIC HEALTH RELEVANCE: In order to better identify and counsel persons with unrecognized HIV infection and link them to clinical and prevention services as well as further reduce perinatal transmission of HIV in the United States the CDC has recommended expanded screening. The most sensitive screening method is based upon detection of the nucleic acid testing but this approach is currently expensive and performed in reference laboratories. We propose the miniaturization and automation of nucleic acid testing methods to enable low cost, near patient testing.