Multiplex Serodiagnostic Assays for Pathogenic Arboviruses in Brazil

巴西致病性虫媒病毒的多重血清诊断检测

• 批准号: 10642843
• 负责人: Wei-Kung Wang
• 金额: $ 15.65万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-05 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10642843
• 关键词: Acute; Age; Alphavirus; Ambulatory Care Facilities; Antibodies; Antigens; Arbovirus Infections; Arboviruses; Binding; Biological Assay; Brazil; Chikungunya virus; Communities; Country; Data; Dengue Infection; Dengue Virus; Diagnosis; E protein; Employment; Enrollment; Enzyme-Linked Immunosorbent Assay; Epidemiology; FDA Emergency Use Authorization; Fever; Flavivirus; Flavivirus Infections; Geographic Locations; Household; IgG3; Immunoassay; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Mayaro virus; Microspheres; Mutate; Mutation; Neutralization Tests; Nonstructural Protein; Nucleocapsid; Nucleocapsid Proteins; Oropouche virus; Orthobunyavirus; Pathogenicity; Patients; Phase; Plasma; Proteins; Publications; Recombinant Proteins; Recombinants; Research; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Sensitivity and Specificity; Serology; Serology test; Seroprevalences; Serotyping; Serum; Site; Testing; Vaccination; Vaccines; Virus Diseases; Virus-like particle; West Nile virus; Yellow Fever; Yellow fever virus; ZIKV infection; Zika Virus; antibody detection; cross reactivity; experience; follow-up; implementation facilitation; innovation; member; recombinant virus; serosurveillance; transmission process

Abstract A critical need exists for highly sensitive and specific serodiagnostic tests to discriminate infections by pathogenic arboviruses in geographic regions, such as Brazil, where multiple flaviviruses including Zika virus (ZIKV), four serotypes of dengue virus (DENV1-4), West Nile virus (WNV), and yellow fever (YFV), as well as chikungunya virus (CHIKV), Mayaro virus (MAYV) (both alphaviruses) and Oropouche virus (OROV) (a bunyavirus), are endemic. However, cross-reactivity of antibodies against different flaviviruses present major challenges, such that even neutralization tests cannot confirm a specific flavivirus infection among individuals who have experienced previous flavivirus infections. The objective of the proposed research is to develop highly sensitive and specific serodiagnostic tests to discriminate infections by pathogenic arboviruses. The central hypothesis is that a combination of fusion loop (FL)-mutated VLPs and nonstructural protein 1 (NS1) proteins of different flaviviruses and recombinant proteins and VLPs of CHIKV, MAYV and OROV in multiplex formats can distinguish different arbovirus infections. The first Aim is to develop and validate multiplex IgG and IgM microsphere immunoassays (MIAs) based on recombinant proteins to distinguish arbovirus infections. We will employ recombinant NS1 proteins of 7 flaviviruses, envelope (E) 2 protein and nucleocapsid (N) protein of CHIKV, MAYV and OROV for multiplex IgG and IgM MIAs using Luminex 200 and test with 15 panels of convalescent-phase serum/plasma from individuals with confirmed arbovirus infections. The second Aim is to develop and validate multiplex IgG and IgM MIAs based on VLPs to distinguish arbovirus infections. We will use purified and FL-mutated VLPs of 7 flaviviruses and VLPs of CHIKV, MAYV and OROV, and test with 15 panels of serum/plasma as in Aim 1. The third Aim is to compare the multiplex IgM MIAs and IgG MIAs with currently available serodiagnostic assays to determine arbovirus seroprevalence in Bahia, a northeastern state with multiple arbovirus transmissions in Brazil. For serodiagnosis, we will test serum samples from patients with acute febrile illness and their follow-up at outpatient clinics of 4 study sites (Salvador, Feira de Santana, Itabuna and Campo Formoso) in Bahia. For seroprevalence study, we will enroll and test household members at communities of the 4 study sites. The proposed study is innovative as it employs two promising antigens (NS1 protein and FL-mutated VLPs) to overcome flavivirus cross-reactivity for seven flaviviruses, as opposed to traditional E protein-based tests, plus CHIKV, MAYV and OROV in two multiplex formats to discriminate arbovirus infections in Brazil. It would contribute to a detailed understanding of the epidemiology of 10 arbovirus infections in Bahia. The successful employment of the multiplex platforms can serve as a new paradigm for serodiagnosis and serosurveillance in countries where multiple arboviruses co-circulate. Most importantly, this research will facilitate the implementation of arbovirus vaccines, in particular ZIKV, DENV and CHIKV vaccines in endemic regions.

抽象的 迫切需要高度敏感和特异性的血清诊断测试来区分感染 巴西等地理区域存在致病性虫媒病毒，该地区存在包括寨卡病毒在内的多种黄病毒 (ZIKV)、登革热病毒 (DENV1-4)、西尼罗河病毒 (WNV) 和黄热病 (YFV) 四种血清型，以及 基孔肯雅病毒 (CHIKV)、马亚罗病毒 (MAYV)（均为甲病毒）和奥罗普切病毒 (OROV)（一种 布尼亚病毒），是地方性的。然而，针对不同黄病毒的抗体的交叉反应性存在重大问题。 挑战，即使是中和测试也无法确认个体中特定的黄病毒感染 曾经感染过黄病毒的人。 拟议研究的目的是开发高度敏感和特异的血清诊断测试 区分致病性虫媒病毒感染。中心假设是融合循环的组合 不同黄病毒的 (FL) 突变 VLP 和非结构蛋白 1 (NS1) 蛋白以及重组蛋白 多重形式的CHIKV、MAYV和OROV VLP可以区分不同的虫媒病毒感染。 第一个目标是开发和验证多重 IgG 和 IgM 微球免疫测定 (MIA) 重组蛋白来区分虫媒病毒感染。我们将使用 7 种重组 NS1 蛋白 黄病毒、CHIKV、MAYV 和 OROV 的包膜 (E) 2 蛋白和核衣壳 (N) 蛋白，用于多重 IgG 使用 Luminex 200 进行 IgM MIA 检测，并使用 15 组来自个体的恢复期血清/血浆进行测试 已确诊虫媒病毒感染。第二个目标是开发和验证基于多重 IgG 和 IgM MIA 在 VLP 上区分虫媒病毒感染。我们将使用 7 种黄病毒和 VLP 的纯化和 FL 突变的 VLP CHIKV、MAYV 和 OROV，并按照目标 1 中的 15 组血清/血浆进行测试。第三个目标是比较 多重 IgM MIA 和 IgG MIA 以及当前可用的血清诊断测定法来确定虫媒病毒 巴西东北部巴伊亚州存在多种虫媒病毒传播。对于血清学诊断， 我们将在 4 项研究的门诊诊所检测急性发热性疾病患者的血清样本及其随访情况 位于巴伊亚的遗址（萨尔瓦多、费拉德桑塔纳、伊塔布纳和坎波福尔莫索）。对于血清流行率研究，我们将 在 4 个研究地点的社区招募并测试家庭成员。 拟议的研究具有创新性，因为它采用了两种有前途的抗原（NS1 蛋白和 FL 突变的 VLP） 与传统的基于 E 蛋白的测试相反，克服了七种黄病毒的黄病毒交叉反应性，加上 CHIKV、MAYV 和 OROV 以两种多重格式来区分巴西的虫媒病毒感染。它会 有助于详细了解巴伊亚州 10 种虫媒病毒感染的流行病学。成功者 多重平台的使用可以作为血清诊断和血清监测的新范例 多种虫媒病毒共同传播的国家。最重要的是，这项研究将促进 在流行地区实施虫媒病毒疫苗，特别是 ZIKV、DENV 和 CHIKV 疫苗。

项目成果

Erratum for Herrera et al., "T Cell Responses to Nonstructural Protein 3 Distinguish Infections by Dengue and Zika Viruses".

Herrera 等人的勘误，“T 细胞对非结构蛋白 3 的反应区分登革热和寨卡病毒感染”。

• 发表时间: 2018-10-23
• 影响因子: 6.4
• 作者: Herrera, Bobby Brooke;Tsai, Wen;Brites, Carlos;Luz, Estela;Pedroso, Celia;Drexler, Jan Feli;Wang, Wei;Kanki, Phyllis J
• 通讯作者: Kanki, Phyllis J

Potential Point-of-Care Testing for Dengue Virus in the Field.

潜在的现场登革热病毒即时检测。

• 发表时间: 2018-05
• 影响因子: 9.4
• 作者: Wang, Wei;Gubler, Duane J
• 通讯作者: Gubler, Duane J

Epidemiology and Immunopathogenesis of Ebola and Flaviviruses

埃博拉病毒和黄病毒的流行病学和免疫发病机制

• DOI: 10.1016/j.coi.2018.05.001
• 发表时间: 2018-05-09
• 期刊: Current opinion in immunology
• 影响因子: 7
• 作者: B. B. Herrera

Potent Neutralizing Human Monoclonal Antibodies Preferentially Target Mature Dengue Virus Particles: Implication for Novel Strategy for Dengue Vaccine.

有效的中和人类单克隆抗体优先针对成熟的登革热病毒颗粒：对登革热疫苗新策略的启示。

• 发表时间: 2018-12-01
• 作者: Tsai, Wen;Chen, Hui;Tsai, Jih;Dejnirattisai, Wanwisa;Jumnainsong, Amonrat;Mongkolsapaya, Juthathip;Screaton, Gavin;Crowe Jr, James E;Wang, Wei
• 通讯作者: Wang, Wei

Dengue outbreaks in Hawai'i After WWII - A Review of Public Health Response and Scientific Literature.

二战后夏威夷爆发登革热 - 公共卫生应对和科学文献回顾。

• 发表时间: 2018-12
• 作者: Lew, Rachel J;Tsai, Wen;Wang, Wei
• 通讯作者: Wang, Wei