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

摘要 寨卡病毒（ZIKV）感染对胎儿的神经发育具有破坏性影响， 南美洲和中美洲的主要健康问题，并可能影响美国部分地区。可靠 ZIKV的诊断工具是至关重要的，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， 怀孕期间的风险管理;评估疫苗和疗法的效力;跟踪 感染，跟踪ZIKV对胎儿发育的影响;协助根除工作;并确保 血液供应安全。目前，根据CDC，没有可靠的ZIKV诊断工具可用， 管道技术主要集中在适合于集中实验室的方法，而不适合于 资源贫乏的环境和护理点。宾夕法尼亚大学和 和巴拿马的戈尔加斯健康研究所（ICGES）将研究和 开发用于ZIKV和ZIKV的组合分子检测的混合的、廉价的、床旁护理系统。 口腔样品中的ZIKV抗体。该系统可用于临床，医生和 牙医办公室，以及在家里用最少的仪器和（可选地）没有电力。减少 该系统利用智能手机进行信号检测、处理， 通讯该系统以其拟议的形式适用于个人诊断，但也适合 同时处理大量样品。宾夕法尼亚大学的研究小组将开发这种检测方法： 病毒核酸的转录等温酶促扩增（RT-LAMP）， 扩增子和免疫测定以分别监测抗体与ZIKV和登革病毒的相互作用 （DENV）抗原以提高测定特异性;以及必要的硬件。ICGES将验证系统的 性能和易用性与ZIKV患者在巴拿马的样本。

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.