Multiplexed Detection of Mosquito-Borne Viruses at the Point-of-Care

在护理点对蚊媒病毒进行多重检测

• 批准号: 10349474
• 负责人: Z. Hugh Fan
• 金额: $ 37.19万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-11 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10349474
• 关键词: Ache; Address; Affect; Americas; Anxiety; Arboviruses; Area; Arthralgia; Bacterial Infections; Benchmarking; Biological Assay; Blood; Blood Circulation; Blood specimen; Brazil; Caribbean region; Centers for Disease Control and Prevention (U.S.); Chikungunya virus; Clinic; Clinical; Clinical Management; Coin; Country; Cytolysis; DNA Primers; Dengue; Dengue Infection; Dengue Shock Syndrome; Dengue Virus; Detection; Devices; Diagnosis; Diagnostic; Disease Outbreaks; Enzyme-Linked Immunosorbent Assay; Epidemic; Exanthema; Fever; Florida; General Population; Genetic; Goals; Haiti; Headache; Hospitals; Human; Human Resources; Infant; Infection; Laboratories; Latin America; Life; Link; Mayaro virus; Mediating; Medical; Methods; Microcephaly; Monitor; Mothers; Neurologic Deficit; Newborn Infant; Nucleic Acids; Paper; Patient Monitoring; Patients; Performance; Polymerase Chain Reaction; Population; Pregnancy; Preparation; Process; Public Health; RNA; RNA Viruses; RNA amplification; Reporting; Research; Reverse Transcription; Saliva; Sampling; Sensitivity and Specificity; Serotyping; Symptoms; Syndrome; Testing; Time; Urine; Virus; Virus Diseases; World Health Organization; ZIKV infection; Zika Virus; amplification device; base; chikungunya infection; clinical care; cost; design; detection platform; detection sensitivity; diagnostic platform; disability; improved; innovation; interest; isothermal amplification; joint inflammation; mosquito-borne; multiplex detection; operation; pathogen; patient screening; point of care; point of care testing; point-of-care diagnostics; population based; saliva sample; screening; vector mosquito; viral detection; viral transmission; virtual

Project Summary Dengue virus (DENV), Zika virus (ZIKV), Chikungunya virus (CHIKV), and Mayaro virus (MAYV) are all mosquito-borne RNA viruses. They are public health concerns because (1) DENV and CHIKV cause hundreds of millions of infections each year, with significant burdens in affected areas, (2) the outbreak of ZIKV in Brazil in 2015/2016 caused anxieties to general population due to its association with microcephaly of newborns, and (3) MAYV emerged in Central and Southern America recently and has the potential for epidemic spread. Because these virus infections have virtually identical clinical presentation and they often circulate concurrently, it is important to have a point-of-care (POC) testing platform to accurately identify virus infection for clinical management of patients, including different complications from these viruses. According to the Centers for Disease Control and Prevention (CDC), the current methods authorized for assessing DENV, ZIKV, and CHIKV infections include reverse transcription polymerase chain reaction (RT- PCR) assay and enzyme-linked immunosorbent assay (ELISA). However, these assays are carried out in laboratories, not at POC in a clinic or in an infected field. It is also important to note that virus infection can cause asymptomatic infections, up to 80% of ZIKV patients, 50% of DENV patients, and 28% of CHIKV patients, respectively. As a result, POC testing in the field will be more valuable for screening asymptomatic patients and monitoring possible virus transmission than a laboratory test because only symptomatic patients go to hospitals or clinics for seeking medical help or to be screened. To address the need, we propose to develop a POC diagnostic platform called Valve-enabled Lysis, paper- based RNA Enrichment, and RNA Amplification Devices (VLEAD). VLEAD will integrate sample preparation— including virus lysis and RNA enrichment—with nucleic acid amplification for simultaneous detection of these viruses. To achieve the goal, we aim to (1) develop multiplexed VLEAD for simultaneous detection of ZIKV, DENV, CHIKV and MAYV; (2) optimize VLEAD using various samples and compare the suitability of the device for urine, saliva and blood samples; and (3) validate VLEAD using clinical samples and compare VLEAD with the benchmark methods including conventional RT-PCR. The significance of the research lies in the following aspects. First, these mosquito-borne RNA viruses are a public health concern. An accurate and sample-to-answer virus detection platform at POC will be useful for clinical care and patient management. Second, a large percentage of these virus infections are asymptomatic, thus a non-invasive POC platform would be very beneficial for screening the general population in the infected area and monitoring virus transmission. Third, the VLEAD platform can be adapted for detecting other pathogens of interest, with a potential to have more societal impacts.

项目摘要 登革热病毒（DENV），寨卡病毒（Zikv），Chikungunya病毒（Chikv）和Mayaro病毒（Mayv）都是 蚊子传播的RNA病毒。它们是公共卫生问题，因为（1）DENV和CHIKV会导致数百个 每年有数百万感染，在受影响地区有明显的烧伤，（2）巴西ZIKV爆发 2015/2016年，由于其与新生儿的小头畸形的关联，使一般人口造成了动画，并且 （3）Mayv最近出现在中美洲和南美，并具有流行病的潜力。 因为这些病毒感染实际上具有相同的临床表现，并且经常圆 同时，重要的是要有一个护理点（POC）测试平台，以准确识别病毒感染 用于患者的临床管理，包括与这些病毒不同的并发症。 根据疾病控制与预防中心（CDC）的说法，当前授权的方法 评估DENV，ZIKV和CHIKV感染包括逆转录聚合酶链反应（RT- PCR）测定和酶联免疫吸附测定法（ELISA）。但是，这些测定是在 实验室，不在诊所或感染领域的POC。同样重要的是要注意病毒感染可以 引起渐近感染，多达80％的ZIKV患者，50％的DENV患者和28％的CHIKV 患者分别。结果，该领域的POC测试对于筛查无症状将更有价值 患者和监测可能的病毒传播比实验室检查，因为只有症状患者 去医院或诊所寻求医疗帮助或筛查。 为了满足需求，我们建议开发一个POC诊断平台，称为启用阀门的裂解，纸张 - 基于RNA的RNA富集和RNA扩增设备（Vlead）。 Vlead将整合样品制备 - 包括病毒裂解和RNA酶 - 以及核酸扩增以简单检测这些 病毒。为了实现目标，我们的目标是（1）开发多重vlead以简单检测ZIKV， Denv，Chikv和Mayv； （2）使用各种样品优化Vlead，并比较设备的适用性 用于尿液，唾液和血液样本； （3）使用临床样本验证Vlead，并将Vlead与 包括常规RT-PCR的基准方法。 研究的意义在于以下方面。首先，这些蚊子传播的RNA病毒是 公共卫生问题。 POC的准确和样本到回答的病毒检测平台将对 临床护理和患者管理。其次，这些病毒感染中很大一部分是不对称的， 因此，非侵入性POC平台将对筛查感染的一般人群非常有益 区域和监测病毒传播。第三，可以调整Vlead平台以检测其他 感兴趣的病原体，有可能产生更多的社会影响。