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Dr
Juliann Makau

Kenya
Kenya

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Profile EDCTP Fellowship 1

Project Title

Determination of HIV-1 drug resistance among patients failing second-line treatment in Kenya using an in-house phenotypic assay (HIVDR)

EDCTP Project

TMA2019CDF-2716

EDCTP Program

EDCTP2

EDCTP Project Call

Career Development Fellowship (CDF)

Project Objectives

- Establish and validate an in-house phenotypic assay for direct isolation of infectious HIV from plasma. -Use the in-house phenotypic assay to describe drug resistance patterns in patients failing second-line treatment in Kenya.

Study Design

Descriptive epidemiology study

Project Summary

Medicines for HIV management have made it possible for people living with HIV to lead a normal life. However, when people fail to take the medicines as recommended by the doctor or they take the medicines for many years, the virus changes in a way that the medicines stop working well. This is a big problem because people will progressively get ill despite being on treatment. It is important to perform tests that determine whether the medicines that someone is taking are working well. In this study, we hoped to develop a new test that will enable the direct observation of how virus from patient blood samples grows in the laboratory in the presence of HIV medicines. We also intended to compare the data generated from the new assay with drug resistance data from sanger sequencing. We recruited study participants from a cohort of children and adolescents living with HIV in eight suburbs of Nairobi.

Host Organisation

Department Institution Country
Centre for Virus Research Kenya Medical Research Institute (KEMRI) KE
Centre for Virus Research Kenya Medical Research Institute (KEMRI) KE

Results & Outcomes

We developed an in-house phenotypic assay for HIV-1 culture by optimizing the conditions required to culture HIV-1 derived from plasma samples in TZM-bI cells, a reporter cell line. We prepared polycationic amyloid fibrils that we used to capture the HIV in samples. We found that the amyloid fibrils enhanced HIV infectivity of a lab strain, HIV-1BaL and HIV-1 from freshly collected blood samples. HIV-1 infectivity was greatly reduced upon freezing and thawing and this made it difficult to perform drug resistance testing for collected blood samples using the phenotypic assay. We thus used genotypic test and found 3 out of 30 individuals had triple class drug resistance and one of them was extensively resistant to all protease inhibitors (PIs), nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). The individual had four mutations in the protease gene including M46I, I47V, I54V, and V82A which are known to confer resistance to all PIs. In addition, NRTI mutations including M41L, M184V, L210W, T215Y and NNRTI mutations including A98G, K103N are known to cause resistance to all NRTIs and NNRTIs, respectively. Our analysis showed that 12 samples exhibited dual class drug resistance mutations to NRTIs and NNRTIs, while 4 samples were resistant to all NRTIs and NNRTIs. These data depict the extent of drug resistant mutations in the sampled cohort of children and adolescents living with HIV and sheds more light on the importance of monitoring drug resistance and proper optimization of treatment regimen for better treatment outcomes.