Abstract |
In this study, we described an ultrasensitive and high-throughput luciferase immunosorbent assay (LISA) for qualitative and quantitative detection of anti-HIV-1 antibody. Anti-HIV antibody in serum or plasma samples was captured by protein A/G-coated microtiter plate and detected with crude cell lysates expressing Nanoluc luciferase (Nluc) enzyme fused with HIV-1 p24 or gp41 antigen without the need of protein purification. After the addition of furimazine substrate, anti- HIV antibodies were quantitatively measured as luciferase light units. LISA showed a wide linear range of detection and was about 104-fold more sensitive than ELISA. For the detection of both anti-p24 and anti-gp41, LISA showed extraordinary sensitivity (99.5% and 100%, respectively) and equivalent specificity (100%). LISA could also monitor the change in the anti-HIV-1 antibody response over time in antiretroviral therapy (ART) treated individuals, and can sufficiently distinguish between recent and long-term HIV-1 infections. Our preliminary results indicate that LISA may provide a novel universal immunoassay platform for simultaneous HIV-1 detection, quantitative measurement of anti- HIV antibodies as well as the differentiation of HIV-1 infection stages.
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Authors | Haiying Wang, Qundi Cai, Yuanhao Liang, Jingwei Shui, Shixing Tang |
Journal | Virus research
(Virus Res)
Vol. 263
Pg. 9-15
(04 02 2019)
ISSN: 1872-7492 [Electronic] Netherlands |
PMID | 30605754
(Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2018. Published by Elsevier B.V. |
Chemical References |
- HIV Antibodies
- HIV Core Protein p24
- HIV Envelope Protein gp41
- gp41 protein, Human immunodeficiency virus 1
- Luciferases
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Topics |
- Diagnostic Tests, Routine
(methods)
- HIV Antibodies
(blood)
- HIV Core Protein p24
(immunology)
- HIV Envelope Protein gp41
(immunology)
- HIV Infections
(diagnosis)
- HIV-1
(immunology)
- Immunosorbent Techniques
- Luciferases
(analysis)
- Luminescent Measurements
(methods)
- Protein Binding
- Sensitivity and Specificity
- Staining and Labeling
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