Molecular and Cellular Biomedical Sciences (MCBS)
Vol 6, No 2 (2022)

Development of Recombinant Immunoblot Assay Diagnostic Test Based on HIV-1 in Indonesia

Jeanne Elvia Christian (Master Program of Biomedical Science, Faculty of Medicine, Universitas Indonesia, Jakarta)
Silvia Tri Widyaningtyas (Virology and Cancer Pathobiology Research Center, Faculty of Medicine, Universitas Indonesia/Cipto Mangunkusumo General Hospital, Jakarta)
Budiman Bela (Virology and Cancer Pathobiology Research Center, Faculty of Medicine, Universitas Indonesia/Cipto Mangunkusumo General Hospital, Jakarta)



Article Info

Publish Date
05 Jul 2022

Abstract

Background: High mutation rates in HIV-1 could affect the accuracy of diagnostic tests. Therefore, recombinant antigen that has an immunodominant and conserved region from HIV-1 need to be developed to detect HIV-1 infection in Indonesia.Materials and methods: The recombinant antigens comprise of Gag (p24), Pol and Env (gp41). Each antigens was expressed in the Escherichia coli expression system and purified using Ni-NTA chromatography. The reactivity of purified antigen against HIV antibodies was tested against a group of 50 HIV-positive plasma samples and 45 HIV-negative plasma samples in a recombinant immunoblot assay (RIBA) platform test. Moreover, 21 of 50 HIV-positive samples and 3 of 45 HIV-negative samples were also tested using HIV blot 2.2 to compare RIBA with a commercial western blot kit. Ten HBV-positive and 10 HCV-positive plasma samples were used to check cross-reactivity with HIV recombinant proteins in RIBA.Results: All HIV-positive samples (100%) tested with RIBA were reactive towards Gag (p24), Pol, Env (gp41). Otherwise, 3 of 21 HIV-positive samples assayed with HIV blot 2.2 were not reactive to Pol protein. All HIV-negative samples tested with RIBA and 3 HIV-negative samples tested with HIV blot 2.2 did not produce any bands of HIV antigens. Few HBV and HCV samples showed reactivity towards HIV recombinant proteins.Conclusion: Each recombinant protein, Gag (p24), Pol, Env (gp41), could be expressed and purified, as well as had reactivity to HIV-positive samples in RIBA test. Therefore, RIBA can be used as a diagnostic test to detect HIV-1 infection in Indonesia.Keywords: diagnostic, HIV-1, immunodominant, recombinant immunoblot assay (RIBA)

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Journal Info

Abbrev

mcbs

Publisher

Subject

Biochemistry, Genetics & Molecular Biology Dentistry Immunology & microbiology Medicine & Pharmacology Neuroscience

Description

Molecular and Cellular Biomedical Sciences (MCBS) has been published by Cell and BioPharmaceutical Institute (CBPI), a biannually published scientific journal, is an open access, peer-reviewed journal that supports all topics in Biology, Pathology, Pharmacology, Biochemistry, Histology and ...