Molecular and Antibody Detection of Zika Virus in Saliva at the Point of Care

摘要 寨卡病毒(ZIKV)感染对胎儿神经发育的破坏性影响已经是一种 南美洲和中美洲的主要健康问题，并可能影响到美国的部分地区。可靠 ZIKV的诊断工具至关重要，因为ZIKV感染者要么没有症状，要么没有特异性 症状类似于其他更常见的病毒感染。这些诊断工具也是必不可少的 孕期风险管理；评估疫苗和疗法的有效性；跟踪病毒传播 感染，追踪寨卡病毒对胎儿发育的影响；协助根除努力；并确保 血液供应安全。目前，根据疾控中心的说法，没有可靠的ZIKV诊断工具可用， 管道技术主要集中在适用于集中式实验室的方法，而不适用于 资源贫乏的环境和关注点。宾夕法尼亚大学的这项合作努力 和巴拿马的研究开发研究所(ICGES)将研究和 开发一种廉价的混合护理点系统，用于寨卡病毒和猪瘟的联合分子检测 口腔样本中的寨卡病毒抗体。该系统可应用于临床、医生和医疗机构 牙医办公室，在家里使用最少的仪器，并且(可选)没有电力。减少 成本，实现量化，该系统利用智能手机进行信号检测、处理和 通讯。该系统，在其建议的形式，适用于个人诊断，但服从 并发处理大量样本。宾夕法尼亚大学的研究小组将开发这种分析方法：反转 目视检测病毒核酸转录等温酶扩增(RT-LAMP) 用扩增和免疫测定法分别监测抗体与寨卡病毒和登革病毒的相互作用 (DENV)抗原，以提高检测的特异性；以及必要的硬件。ICGES将验证系统的 巴拿马ZIKV患者样本的性能和易用性。

ABSTRACT Zika virus (ZIKV) infection with its devastating impact on neurological development of fetuses is already a major health problem in South and Central America, and is likely to impact parts of the USA. Reliable diagnostic tools for ZIKV are vital since ZIKV-infected individuals display either no symptoms or nonspecific symptoms similar to other, more common, viral infections. These diagnostic tools are also essential for management of risks during pregnancy; assessing efficacy of vaccines and therapies; tracking the spread of the infection, tracking ZIKV effects on fetal development; assisting in eradication efforts; and assuring the safety of blood supply. Currently, according to the CDC, no reliable diagnostic tools for ZIKV are available and pipeline technologies are mostly focused on methods suitable for centralized laboratories and inappropriate for resource poor settings and the point of care. This collaborative effort between the University of Pennsylvania and the Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES) in Panama will research and develop a hybrid, inexpensive, point of care system for combined molecular detection of ZIKV and of antibodies against the ZIKV in oral samples. The proposed system can be used in the clinic, doctor and dentist office, and at home with minimal instrumentation and (optionally) without electrical power. To reduce cost, enable quantification, the system utilizes a smartphone for signal detection, processing, and communications. The system, in its proposed form, is suitable for individual diagnostics, but is amenable to concurrent processing of large numbers of samples. The Penn team will develop the assays: reverse transcription isothermal enzymatic amplification (RT-LAMP) of viral nucleic acids with visual detection of amplicons and an immunoassay to monitor separately antibodies interactions with ZIKV and dengue virus (DENV) antigens to improve assay specificity; and the necessary hardware. ICGES will verify system's performance and ease of use with samples of ZIKV patients in Panama.