Photonic Crystal Based Point-of-Care Detection of HIV Viral Load Using a Smartphone Biosensor

 描述（由申请人提供）：艾滋病毒/艾滋病流行病是当代社会最大的全球健康挑战之一，对年轻妇女产生了严重影响，特别是在世界上许多最贫穷的地区。由于2500万人死亡，另有3500万人感染，预防，诊断和治疗是任何遏制疾病计划的关键组成部分。虽然没有治愈方法，但抗逆转录病毒疗法（ART）已被证明可以成功地减缓疾病的进展，方法是在患者体内保持低数量的病毒，使他们的CD 4计数保持在可以成功抵御机会性感染的水平。世界卫生组织（WHO）已将病毒载量（VL）检测确定为监测接受抗逆转录病毒治疗患者的首选方法;然而，目前的金标准技术依赖于核酸检测，这依赖于昂贵的、技术上具有挑战性的设备，需要稳定的电力和温度控制等基础设施。由于无法在受该疾病影响最严重的许多地区进行VL检测，医院和诊所不得不依赖不太准确的诊断。该提案旨在开发一种即时检测解决方案，以满足世界各地无法获得VL检测的临床医生和患者的需求。拟议的解决方案将使用连接到光子晶体的微流体芯片来检测100微升患者血液样本中的病毒颗粒数量，该光子晶体由连接到标准智能手机的定制光学支架读取，并且灵敏度至少为200拷贝/mL，比WHO建立的ART监测阈值低5倍。为了实现这一目标，将根据工作概念验证原型设计自参考光学支架，然后使用HIV-1 gp 120和抗gp 120之间的相互作用对光谱分辨率和生物附着灵敏度进行表征。微流控芯片将被设计在光子晶体（PC）生物传感器的顶部，以减少工作体积，提高灵敏度，并增加易用性。然后，将对加标患者血浆样本进行检测，以量化器械的检测限（LOD）。通过基于电流体动力学喷射印刷和基于折射率的信号放大分子的制造工艺进一步使微流体装置微化，将实现LOD的进一步改进。最后，全血样本将用于确定新的LOD，HIV阳性和阴性患者的样本将用于验证系统的实用性。

 DESCRIPTION (provided by applicant): The HIV/AIDS pandemic is one of the single greatest global health challenges of contemporary society that disproportionally affects young women, especially in many of the world's poorest regions. With 25 million dead and another 35 million infected, prevention, diagnosis, and treatment are all crucial components of any plan to curb the disease. While no cure exists, antiretroviral therapies (ART) have proven successful in slowing down the progression of the disease by maintaining low numbers of virus inside patients so that their CD4 counts are maintained at levels which can successfully fight off opportunistic infections. The World Health Organization (WHO) has identified Viral Load (VL) testing as the preferred method to monitor patients on ART; however, current gold-standard techniques rely on nucleic-acid testing which are dependent on expensive, technically challenging equipment with a requirement for infrastructure such as stable electricity and temperature control. The inability to perform VL testing in many of the areas most heavily affected by the disease has left hospitals and clinics to rely on less-accurate diagnostics. This proposal seeks to develop a point of care testing solution that will address the needs of clinicians and patients across the world who do not have access to VL testing. The proposed solution will sense the number of virus particles in a 100 µL sample of patient blood using a microfluidic chip attached to a photonic crystal which is read by a custom optical cradle attached to a standard smartphone and sensitive down to at least 200 copies/mL, 5x below the WHO established threshold for ART monitoring. To accomplish this, a self-referencing optical cradle will be designed based upon working proof-of-concept prototypes that will then be characterized for both spectral resolution and sensitivity to biological attachment using the interactions between HIV-1 gp120 and anti-gp120. A microfluidic chip will be designed on top of a photonic crystal (PC) biosensor to decrease the working volumes, increase sensitivity, and increase ease-of-use. This system will then be tested on spiked patient plasma samples to quantify a limit of detection (LOD) for the device. Further improvements of the LOD will be achieved by further miniaturizing the microfluidics device through a manufacturing process based upon electrohydrodynamic-jet printing, and refractive-index based signal amplifying molecules. Finally, whole blood samples will be used to determine a new LOD and samples from HIV positive and negative patients will be used to validate the utility of the system.