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）和丙型肝炎病毒（HBV）共享相同的血液模式 传播，主要是通过性接触和注射药物使用和有艾滋病毒风险的人的传播是 HBV感染的风险更高。在全球范围内，估计有2.57亿人患有慢性HBV感染 （2015年），而在2019年，有3800万人患有艾滋病毒（PLHIV）。非洲，特别是撒哈拉次 非洲（SSA）是世界上受影响最大的地区，PLHIV为70％（2570万），约占6％ 感染HBV的成年人口。重要的是，据估计，慢性HBV感染影响了5-20％ plhiv。 HBV是一种不对称的肝病，使其早期发现困难并导致严重 肝硬化，肝细胞癌和早期死亡等并发症。了解HBV状态 HIV抗逆转录病毒疗法（ART）的启动对于适当选择初始艺术方案至关重要 替诺福韦毒素富马酸（TDF）应与拉米夫丁（3TC）或Emtricitabine（FTC）相结合， 抑制了HIV和HBV复制，并用于监测治疗。马里，西非的LMIC 艾滋病毒/艾滋病患病率> 1％的普通人群（> 2000万）和特定的成年人口 组（例如孕妇，学生，献血者）的HBS AG正率从10到 18.8％。虽然建议对HIV RNA和HBV DNA定量测定法进行更好的指导和监测 治疗，在SSA中获得此类量化评估的访问非常有限，甚至没有可用。在大多数 非洲，HIV患者仅在HBS AG上进行筛查。那是隐匿性丙型肝炎的患者（OBI），多pite 血清中的HBV-DNA和/或肝脏中的HBV-DNA，但HBS Ag谈判将不会被检测到，因此错过了重要的 启动治疗的机会。 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，例如受艾滋病毒感染的人。病毒核酸的测量 在确定感染阶段，选择治疗方面的关键作用，并了解效率 抗病毒疗法。该K43应用程序旨在开发多重，实时，定量PCR（RT- QPCR）用于简单定量HIV RNA和HBV DNA的测定，该测定专门设计为 使用经济，环境和 在SSA的实验室人员的技术能力中。我提出的职业发展将是 在美国和马里的导师的监督下，专注于获得设计的专业知识和技能 以及开发新的多重PCR分析，评估新诊断测试的方法以及 针对新测试的实施策略，特别是与利益相关者的参与有关，以及 增强我在研究领导力和团队科学方面的职业技能。