Development of immunoassays for detection of Human Immunodeficiency Virus based on Consensus env gp41 Immunodominant Region Peptide from HIV-1 infections in Kenya

James Kimotho, Zipporah Ng’ang’a, Edna Nyairo, Missiani Ochwoto, Nicholas Nzioka, Francis Ogolla, Michael Kiptoo

Abstract


Background: Human Immunodeficiency Virus (HIV) is characterized by high rates of genetic variability in vivo that could affect the performance of the HIV antibody-based detection kits.

Objective: This study aimed at developing immunoassays for HIV based on Consensus env gp41 Immunodominant region (IDR) from HIV infections in Kenya.

Methods: HIV RNA was extracted from 91 samples collected from 5 regional blood transfusion centers in Kenya. The RNA was reverse transcribed, sequenced in the env gp41-Immunodominant Region (IDR) and the Consensus sequence generated used to synthesize corresponding peptide. The Global HIV envgp41-IDR Consensus peptide was obtained from the literature and also synthesized. The two peptides were used to separately develop HIV immunoassays based on Enzyme-linked Immunosorbent Assay (ELISA) and Lateral Flow Assay (LFA) platforms and the performance of developed assays was evaluated. The same HIV env gp41 IDR peptides were used to develop ELISA-based immunoassays for determination HIV Incidence / Recency.

Results: The study did not find significant difference between the performance of the immunoassays that were developed with Consensus env gp41-IDR peptide (Kenya) and those developed using Consensus env gp41-IDR peptide (Global). However, the study found a significant difference between the performance of HIV ELISA for HIV Incidence testing that was developed with Consensus envgp41-IDR peptide (Kenya) and that which was developed using Consensus envgp41-IDR peptide (Global) with the former displaying superior performance.

Conclusions: The developed immunoassays demonstrated that both Consensus env gp41-IDR peptides (Kenya and Global) could be used to develop HIV immunoassays but Consensus env gp41-IDR peptide (Kenya) could be more suitable for development of HIV Incidence assays in Kenya.

Keywords: HIV, Consensus sequence, env gp41-Immunodominant Region, Immunoassays


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