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: 10295972
• 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/10295972
• 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 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.

总结 人类免疫缺陷病毒（HIV）和B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毒品传播的艾滋病毒感染者和有感染艾滋病毒风险的个人也是 HBV感染的风险更高。全球估计有2.57亿人患有慢性HBV感染 （2015年），而在2019年，有3800万人感染艾滋病毒（PLHIV）。非洲，特别是撒哈拉以南非洲 非洲（SSA）是世界上受影响最严重的地区，有70%（2570万）的艾滋病毒感染者和6%的 成人HBV感染者。重要的是，据估计，慢性HBV感染影响5-20%的 艾滋病毒感染者。HBV是一种无症状的肝脏疾病，使其早期检测困难，并导致严重的 并发症如肝硬化、肝细胞癌和早期死亡。HBV状态的知识 启动HIV抗逆转录病毒治疗（ART）对于适当选择初始ART方案至关重要， 富马酸替诺福韦酯（TDF）应与拉米夫定（3 TC）或恩曲他滨（FTC）联合使用， 其抑制HIV和HBV复制，并用于监测治疗。马里，西非的一个低收入国家 艾滋病毒/艾滋病流行率在总人口（> 2 000万）和特定成人人口中的比例>1% 组（例如，孕妇、学生、献血员）的HBsAg阳性率为10 - 20%。 百分之十八点八虽然HIV RNA和HBV DNA定量检测被推荐用于更好地指导和监测 然而，在SSA中获得这种定量测定的机会非常有限或甚至不可用。大部 在非洲，艾滋病患者只筛查乙肝表面抗原。因此，隐匿性B型肝炎（OBI）患者，尽管有 血清和/或肝脏中的HBV-DNA，但HBsAg阴性，将无法检测到，因此，错过了重要的 有机会开始治疗。OBI通常无症状，但其再激活通常发生在 免疫抑制个体，如HIV感染者。病毒核酸的测定 在确定感染阶段，选择治疗方面起着关键作用，并提供有关疗效的信息 抗病毒治疗。该K43应用寻求开发多重、真实的时间、定量PCR（RT-PCR）。 用于同时定量HIV RNA和HBV DNA的qPCR）检测试剂盒，专门设计用于 检测区域遗传变异，使用一个“开放”的系统平台，这是经济，环境， 在SSA实验室工作人员的技术能力范围内。我建议的职业发展将是 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检测，评估新的诊断测试的方法， 新测试的实施策略，特别是与利益相关者参与有关的策略，以及， 加强我在研究领导和团队科学方面的职业技能。