Improved Diagnosis of Kaposi's Sarcoma-Associated Herpesvirus Infection

改进卡波西肉瘤相关疱疹病毒感染的诊断

• 批准号: 9346755
• 负责人: Enrique A Mesri
• 金额: $ 100万
• 依托单位: EPIPHANY BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-04 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9346755
• 关键词: Acquired Immunodeficiency Syndrome; Age; Algorithms; Antibodies; Antigens; Area; Biological Assay; Biomedical Research; Blinded; Blood; Blood donor; Blood specimen; CD4 Lymphocyte Count; Centers for Disease Control and Prevention (U.S.); Chemoprevention; Clinical; Clinical Research; Code; Collaborations; Consensus; Cyclic GMP; DNA; Data; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Effectiveness; Enzyme-Linked Immunosorbent Assay; Epidemiology; FDA approved; Goals; HIV; HIV Seropositivity; HIV antiretroviral; HIV-1; Health; Herpesviridae; Herpesviridae Infections; High Prevalence; Highly Active Antiretroviral Therapy; Homosexuals; Hospitals; Human Herpesvirus 8; Immune system; Incidence; Individual; Intention; Intervention; Kaposi Sarcoma; Laboratory Research; Life; Life Expectancy; Lytic; Lytic Virus; Malignant Neoplasms; Monitor; Oncornaviruses; Outcome; Patient risk; Patients; Performance; Persons; Phase; Plasma; Population; Preparation; Prevalence; Process; Production; Reagent; Receiver Operating Characteristics; Reproducibility; Research; Research Personnel; Risk; Sampling; Sensitivity and Specificity; Serological; Serotyping; Serum; Signal Transduction; Small Business Innovation Research Grant; Specificity; Testing; Time; Tissue Donors; Tissues; Transplant Recipients; Transplantation; Universities; Variant; Viral Load result; Viremia; Virus Diseases; Western Blotting; Work; analytical method; antiretroviral therapy; base; clinical diagnostics; co-infection; cohort; cost effective; diagnostic assay; field study; healthy volunteer; high risk population; immunosuppressed; improved; large scale production; medical schools; men; method development; prevent; programs; repository; screening; standard measure; synthetic peptide; transmission process; validation studies; viral cyclin; viral transmission

Project Summary/Abstract Kaposi's sarcoma herpes virus (KSHV) is an oncovirus that causes several cancers including Kaposi's sarcoma (KS). It is estimated that 3 to 5% of U.S. blood donors have been infected with KSHV, yet in some high-risk populations such as homosexual men, prevalence is as high as 65%. In spite of HAART, KS remains the second most common AIDS- associated malignancy. A third of AIDS-KS cases now arise in individuals with relatively high CD4+ cell counts and low HIV-1 viral load. A recent study has observed a continuing high prevalence and increased acquisition of KSHV in HIV- 1+ infected persons on HAART. The data clearly indicate that KS is still occurring in HIV-infected individuals, even in the context of successful HAART. In immunosuppressed populations, approximately 1 in 200 transplant patients will develop KS. The prevalence of KSHV is responsible for the continuing incidence of KS and its associated reduction in life expectancy of HIV+ individuals. Despite its strong disease association, particularly in the HIV+ and immunosuppressed populations, there is no FDA approved clinical diagnostic test for KSHV and consequently a significant unmet need remains for a simple, cost-effective commercially viable KSHV diagnostic kit that would be an essential component of any effort to control virus transmission and monitor serostatus of those already infected. The ability to gauge the KSHV serostatus of an HIV+ individual would help clinicians make appropriate interventional choices to prevent KS onset. The overall goal of this project is to develop a serological multi-antigen assay for the detection of KSHV antibodies that can be used commercially to diagnose at-risk patients and identify blood/tissue donors with KSHV infection. In prior SBIR Phase I and II work, Epiphany developed a single-well, multi-antigenic KSHV enzyme linked immunosorbent assay (ELISA) called “4-Combo.” In screening a sera panel containing both KSHV true- positives and normal, low-risk blood donors, 4-Combo was the only assay out of 7 other independent ELISAs to detect all consensus KSHV samples with excellent specificity and sensitivity (estimated >99%, 98%, respectively). For the next stage of the kit development, it is critical to examine 4-Combo's performance against large sera banks of epidemiologically validated KSHV positive and negative controls with relevant confirmatory assays to establish the true effectiveness of the assay. A collaborative consortium that merges Epiphany with the biomedical and clinical research capabilities of the University of Miami (UofM) Miller School of Medicine, the Miami Center for AIDS Research (CFAR) and the labs of Dr. Enrique A. Mesri (CFAR) has been created. Together, Epiphany and Miami CFAR are uniquely equipped to achieve the following Aims. In Aim 1, synthetic, manufacturing, and analytical process development will be implemented followed by pilot production of a set of 4-Combo diagnostic ELISA kits. In Aim 2, these kits will be assessed and optimized by screening retrospective clinical blood samples and serologically characterized plasma/ PMBCs and validated by western blot. In Aim 3, large-scale kit production will commence. Assay stability and validation studies will be performed. A larger panel of HIV+ sera and tissues available to UofM CFAR, its cores and transplant centers, and national repositories will be massively screened to establish sensitivity and specificity. Achieving the goals of this project will establish the clinical and commercial value of 4-Combo ultimately resulting in the first FDA-approved clinical diagnostic assay for KSHV.

项目摘要/摘要 Kaposi的肉瘤疱疹病毒（KSHV）是一种导致包括Kaposi肉瘤（KS）在内的几种癌症的癌病毒。这是 据估计，有3至5％的美国献血者感染了KSHV，但在一些高风险人群中，例如 同性恋者，患病率高达65％。尽管哈特（Haart）仍然是第二个常见的艾滋病 - 相关的恶性肿瘤。目前，CD4+细胞计数较高且低的个体中，三分之一的艾滋病案件出现 HIV-1病毒负荷。最近的一项研究观察到持续的高流行率，并增加了KSHV的HIV- Haart上有1多名感染者。数据清楚地表明，即使在 成功的Haart的背景。在免疫抑制人群中，大约200名移植患者中有1个 开发KS。 KSHV的患病率负责KS的持续事件及其相关的减少 艾滋病毒+个人的预期寿命。尽管疾病的关联很强，尤其是在艾滋病毒+和 免疫抑制人群，没有FDA批准的KSHV临床诊断测试，因此 对于一个简单，具有成本效益的商业可行的KSHV诊断套件的重要需求仍然是 控制病毒传播和监测已经感染的人的血清的任何努力的重要组成部分。这 能够衡量艾滋病毒+个体的KSHV血清表达 选择防止KS发作的选择。该项目的总体目标是开发一种血清学多抗原测定法 检测可在商业上用于诊断高危患者并鉴定血/组织供体的KSHV抗体 带有KSHV感染。在先前的SBIR第一阶段和II工作中，Epiphany开发了单孔多抗原KSHV 酶连接的免疫吸附测定法（ELISA）称为“ 4-Combo”。在筛选一个包含两个kshv true-的血清面板时 阳性和正常的低风险献血者，4-Combo是其他7种独立ELISA中唯一检测所有的测定法 共识KSHV样品具有出色的特异性和灵敏度（估计> 99％，98％）。下一个 套件开发的阶段，考试4-Combo对大型血清库的表现至关重要 通过相关确认测定法，在流行病学上验证的KSHV阳性和负面对照，以确定真实 测定的有效性。一个合作财团，将顿悟与生物医学和临床研究合并 迈阿密大学（UOFM）米勒医学院的能力，迈阿密艾滋病研究中心（CFAR） 以及Enrique A. Mesri博士（CFAR）的实验室。顿悟和迈阿密CFAR在一起是独特的 配备有以下目标。在AIM 1中，合成，制造和分析过程开发将是 随后实施了一套4-Combo诊断ELISA套件的试点生产。在AIM 2中，这些套件将是 通过筛选回顾性临床血液样本和血清学表征血浆/ PMBC进行评估和优化 并通过Western blot验证。在AIM 3中，将开始大规模的套件生产。测定稳定性和验证研究 将执行。 UOFM CFAR，其核心和移植中心以及 国家存储库将进行大规模筛选以建立灵敏度和特异性。实现该项目的目标 将建立4-Combo的临床和商业价值最终导致第一个FDA批准的临床 KSHV的诊断测定法。