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

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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.