Development of a Multiplex Quantitative PCR Assay for HIV and Hepatitis B Virus, for Low- and middle Income Countries

为低收入和中等收入国家开发艾滋病毒和乙型肝炎病毒多重定量 PCR 检测方法

• 批准号: 10472663
• 负责人: DJENEBA BOCAR FOFANA
• 金额: $ 9.96万
• 依托单位: UNIV OF SCIENCES, TECH & TECH OF BAMAKO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-23 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10472663
• 关键词: AIDS/HIV problem; Adult; Affect; Africa; Africa South of the Sahara; African; American; Antiviral Therapy; Antiviral resistance; Award; Biological Assay; Biological Testing; Biostatistical Methods; Blood; Blood donor; Budgets; Burkina Faso; Caring; Cessation of life; Child; Chronic Hepatitis B; Cirrhosis; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Coronavirus; Counseling; Country; Data; Development; Diagnosis; Diagnostic tests; Disease; Doctor of Pharmacy; Doctor of Philosophy; Drug resistance; Early Diagnosis; Early treatment; Electricity; Enterovirus; Environment; Equipment and Supplies; Evaluation; Face; Fumarates; General Population; Genetic Variation; Genotype; Goals; HIV; HIV Infections; HIV antiretroviral; HIV-1; Health Sciences; Health Services Accessibility; Healthcare; Hepatitis B; Hepatitis B Infection; Hepatitis B Surface Antigens; Hepatitis B Virus; Hepatitis C virus; Human Papillomavirus; Human immunodeficiency virus test; Individual; Infection; Knowledge; Laboratories; Laboratory Technicians; Lamivudine; Lead; Leadership; Link; Liver; Liver Cirrhosis; Liver diseases; Mali; Measurement; Measures; Medical; Medical Students; Mentors; Methods; Microbiology; Molecular; Molecular Biology; Monitor; Nucleic Acids; Paris, France; Patient Monitoring; Patients; Performance; Persons; Phase; Play; Population; Population Group; Pregnant Women; Prevalence; Primary carcinoma of the liver cells; Professional Competence; Protocols documentation; Public Health; RNA; Reagent; Refrigeration; Regimen; Research; Resources; Risk; Role; Science; Serum; Source; Students; Supervision; System; Techniques; Technology; Tenofovir; Testing; Time; Training; Universities; Viral; Viral Load result; Virus Diseases; Virus Replication; World Health Organization; animation; anti-viral efficacy; antiretroviral therapy; career development; co-infection; cost; course development; design; emtricitabine; genetic variant; high risk; immunosuppressed; implementation strategy; injection drug use; low and middle-income countries; mid-career faculty; mortality; novel diagnostics; nucleic acid detection; patient screening; population based; programs; research clinical testing; screening; skill acquisition; skills; testing access; tool; tool development; transmission process; viral DNA; virology

SUMMARY Human immunodeficiency virus (HIV) and Hepatitis B virus (HBV) share the same blood borne modes of transmission, primarily through sexual contact and injection drug use and individuals at risk for HIV are also at higher risk for HBV infection. Globally, an estimated 257 million people live with chronic HBV infection (2015) while, in 2019, 38 million people were living with HIV (PLHIV). Africa, in particular sub-Saharan Africa (SSA), is the most affected region in the world with and 70% (25.7 million) of PLHIV and ~6% of the adult population infected with HBV. Importantly, it is estimated that chronic HBV infection affects 5-20% of PLHIV. HBV is an asymptomatic liver disease making its early detection difficult and leading to serious complications such as cirrhosis, hepatocellular carcinoma, and early death. Knowledge of HBV status at initiation of HIV antiretroviral therapy (ART) is essential for appropriate select an initial ART regimen, as tenofovir disoproxil fumarate (TDF) should be combined with lamivudine (3TC) or emtricitabine (FTC), which suppress both HIV and HBV replication, and for monitoring treatment. Mali, a LMIC in West Africa has a HIV/AIDS prevalence rate of >1% of the general population (>20 million) and specific adult population groups (e.g., pregnant women, students, blood donors) have HBs Ag positive rates ranging from 10 to 18.8%. While HIV RNA and HBV DNA quantification assays are recommended to better guide and monitor treatment, access to such quantification assays in SSA is very limited or not even available. In most of Africa, HIV patients screened only on HBs Ag. Thus, patients with occult hepatitis B (OBI), despite having HBV-DNA in serum and/or in liver, but HBs Ag negativity, will not be detected and, thus, miss an important opportunity to initiate treatment. OBI is usually asymptomatic but its reactivation commonly occurs in immunosuppressed individuals, such as in HIV infected persons. Measurement of viral nucleic acids plays a critical role in determining the phase of infection, selecting treatment, and is informative about the efficacy of antiviral therapy. This K43 application seeks to develop a multiplex, real time, quantitative PCR (RT- qPCR) assay for simultaneous quantification of HIV RNA and HBV DNA that is specifically designed to detect regional genetic variants, using an “open” system platform that is economically, environmentally, and within the technical capabilities of laboratory staff in SSA. My proposed career development will be supervised by both American and Malian mentors and focus on gaining expertise and skills in the design and development of new multiplexed PCR assays, methods to evaluate new diagnostic tests, and in implementation strategies for new tests, specifically as it pertains to stakeholder engagement, as well as, strengthen my career skills in research leadership and team science.

概括 人类免疫缺陷病毒（HIV）和乙型肝炎病毒（HBV）具有相同的血液传播模式 主要通过性接触和注射吸毒传播，以及有感染艾滋病毒风险的个人 乙肝病毒感染风险较高 全球估计有 2.57 亿人患有慢性乙肝病毒感染。 （2015）而 2019 年，非洲，特别是撒哈拉以南地区，有 3800 万人感染艾滋病毒（PLHIV）。 非洲 (SSA) 是世界上受影响最严重的地区，有 70%（2570 万）艾滋病毒感染者和约 6% 的感染者 重要的是，感染 HBV 的成年人估计有 5-20% 患有慢性 HBV 感染。 HBV 感染者是一种无症状的肝脏疾病，因此难以早期发现并导致严重的后果。 肝硬化、肝细胞癌和早期死亡等并发症。 开始 HIV 抗逆转录病毒治疗 (ART) 对于适当选择初始 ART 治疗方案至关重要，因为 富马酸替诺福韦二吡呋酯 (TDF) 应与拉米夫定 (3TC) 或恩曲他滨 (FTC) 联合使用， 抑制 HIV 和 HBV 复制，并用于监测西非中低收入国家马里的治疗情况。 总人口（>2000万）和特定成年人口的艾滋病毒/艾滋病患病率>1% 群体（例如孕妇、学生、献血者）的 HBs Ag 阳性率范围为 10 至 18.8%，建议进行 HIV RNA 和 HBV DNA 定量检测以更好地指导和监测。 在 SSA 治疗中，获得此类定量的机会非常有限，甚至在大多数检测中都无法获得。 非洲的 HIV 患者仅接受 HBs Ag 筛查，因此，尽管患有隐匿性乙型肝炎 (OBI)，但患者仍患有隐匿性乙型肝炎 (OBI)。 血清和/或肝脏中的 HBV-DNA，但 HBs Ag 阴性，将不会被检测到，因此会错过一个重要的 OBI 通常是无症状的，但其重新激活通常发生在 免疫抑制的个体，例如艾滋病毒感染者，可以进行病毒核酸的测量。 在确定感染阶段、选择治疗方法方面发挥着关键作用，并且可以提供有关疗效的信息 该 K43 应用旨在开发多重、实时、定量 PCR (RT- qPCR）同时定量 HIV RNA 和 HBV DNA，专门设计用于 使用经济、环保、经济的“开放”系统平台检测区域遗传变异 我建议的职业发展将在 SSA 实验室人员的技术能力范围内。 由美国和马里导师监督，专注于获得设计方面的专业知识和技能 和开发新的多重 PCR 测定、评估新诊断测试的方法以及 新测试的实施策略，特别是与利益相关者参与相关的策略，以及， 加强我在研究领导力和团队科学方面的职业技能。