Summary
HIV-2 infection is confined mostly to West Africa. Seven HIV-2 subtypes have so far been described; only HIV-2 subtypes A and B are prevalent, the others being considered self-limiting infections at the epidemiological level. The main limitation for the HIV-2 DNA proviral quantification is the lack of HIV-2 DNA standard. We designed and tested a new HIV-2 primer couple that amplifies both the HIV-2 ROD strain and HIV-1 LAV/BRU strain. These HIV-2 primers were used to quantified an HIV-2 standard comparatively to a standard widely used in proviral DNA HIV-1 quantification, i.e., the 8E5 cell line transfected by a single defective integrated provirus of HIV-1 BRU/LAV by cell. The primers and probe used to quantify HIV-2 DNA are located in a long terminal repeat (LTR) region with low variability. These primers amplify both HIV-2 subtypes A and B. The relevance of the follow-up of the infected patients by the quantification of the proviral DNA HIV-2 is currently studied.
Key Words
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Clavel, F., Guetard, D., Brun-Vezinet, F., Chamaret, S., Rey, M. A., Santos-Ferreira, M. O., et al. (1986) Isolation of a new human retrovirus from West African patients with AIDS. Science 233, 343–346.
Chen, Z., Luckay, A., Sodora, D. L., Telfer, P., Reed, P., Gettie, A., et al. (1997) Human immunodeficiency virus type 2 (HIV-2) seroprevalence and characterization of a distinct HIV-2 genetic subtype from the natural range of simian immunodeficiency virus-infected sooty mangabeys. J. Virol. 71, 3953–3960.
Gao, F., Yue, L., Robertson, D. L., Hill, S. C., Hui, H., Biggar, R. J., et al. (1994) Genetic diversity of human immunodeficiency virus type 2: evidence for distinct sequence subtypes with differences in virus biology. J. Virol. 68, 7433–7447.
Yamaguchi, J., Devare, S. G., and Brennan, C. A. (2000) Identification of a new HIV-2 subtype based on phylogenetic analysis of full-length genomic sequence. AIDS Res. Hum. Retroviruses 16, 925–930.
De Cock, K. M. and Brun-Vezinet, F. (1989) Epidemiology of HIV-2 infection. AIDS 3(Suppl 1), S89–S95.
Schechter, M. T., Neumann, P. W., Weaver, M. S., Montaner, J. S., Cassol, S. A., Le, T. N., et al. (1991) Low HIV-1 proviral DNA burden detected by negative polymerase chain reaction in seropositive individuals correlates with slower disease progression. AIDS 5, 373–379.
Berry, N., Ariyoshi, K., Jobe, O., Ngum, P. T., Corrah, T., Wilkins, A., et al. (1994) HIV type 2 proviral load measured by quantitative polymerase chain reaction correlates with CD4+ lymphopenia in HIV type 2-infected individuals. AIDS Res. Hum. Retroviruses 10, 1031–1037.
Norrgren, H., Marquina, S., Leitner, T., Aaby, P., Melbye, M., Poulsen, A. G., et al. (1997) HIV-2 genetic variation and DNA load in asymptomatic carriers and AIDS cases in Guinea-Bissau. J. Acquir. Immune Defic. Syndr. Hum. Retrovirol 16, 31–38.
Sarr, A. D., Popper, S., Thior, I., Hamel, D. J., Sankale, J. L., Siby, T., et al. (1999) Relation between HIV-2 proviral load and CD4+ lymphocyte count differs in monotypic and dual HIV infections. J. Hum. Virol. 2, 45–51.
Gomes, P., Taveira, N. C., Pereira, J. M., Antunes, F., Ferreira, M. O., and Lourenco, M. H. (1999) Quantitation of human immunodeficiency virus type 2 DNA in peripheral blood mononuclear cells by using a quantitative-competitive PCR assay. J. Clin. Microbiol. 37, 453–456.
Popper, S. J., Sarr, A. D., Gueye-Ndiaye, A., Mboup, S., Essex, M. E., and Kanki, P. J. (2000) Low plasma human immunodeficiency virus type 2 viral load is independent of proviral load: low virus production in vivo. J. Virol. 74, 1554–1557.
Kanki, P. J., Travers, K. U., Mboup, S., Hsieh, C. C., Marlink, R. G., Gueye, N. A., et al. (1994) Slower heterosexual spread of HIV-2 than HIV-1. Lancet 343, 943–946.
Marlink, R. G., Ricard, D., M’Boup, S., Kanki, P. J., Romet-Lemonne, J. L., N’Doye, I., et al. (1988) Clinical, hematologic, and immunologic cross-sectional evaluation of individuals exposed to human immunodeficiency virus type-2 (HIV-2). AIDS Res. Hum. Retroviruses 4, 137–148.
Damond, F., Gueudin, M., Pueyo, S., Farfara, I., Robertson, D. L., Descamps, D., et al. (2002) Plasma RNA viral load in human immunodeficiency virus type 2 subtype A and subtype B infections. J. Clin. Microbiol. 40, 3654–3659.
Popper, S. J., Sarr, A. D., Travers, K. U., Gueye-Ndiaye, A., Mboup, S., Essex, M. E., and Kanki, P. J. (1999) Lower human immunodeficiency virus (HIV) type 2 viral load reflects the difference in pathogenicity of HIV-1 and HIV-2. J. Infect. Dis. 180, 1116–1121.
Damond, F., Descamps, D., Farfara, I., Telles, J. N., Puyeo, S., Campa, P., et al. (2001) Quantification of proviral load of human immunodeficiency virus type 2 subtypes A and B using real-time PCR. J. Clin. Microbiol. 39, 4264–4268.
Simon, F., Matheron, S., Tamalet, C., Loussert-Ajaka, I., Bartczak, S., Pepin, J. M., et al. (1993) Cellular and plasma viral load in patients infected with HIV-2. AIDS 7, 1411–1417.
Laurendeau, I., Bahuau, M., Vodovar, N., Larramendy, C., Olivi, M., Bieche, I., et al. (1999) TaqMan PCR-based gene dosage assay for predictive testing in individuals from a cancer family with INK4 locus haploinsufficiency. Clin. Chem. 45, 982–986.
Acknowledgments
We thank all the technicians of the Virology Department at the University Hospital Charles Nicolle in Rouen, France. This work was supported by ANRS (Agence Nationale de Recherche sur le SIDA).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Gueudin, M., Damond, F., Simon, F. (2005). Quantification of Proviral DNA Load of Human Immunodeficiency Virus Type 2 Subtypes A and B Using Real-Time PCR. In: Zhu, T. (eds) Human Retrovirus Protocols. Methods in Molecular Biology™, vol 304. Humana Press. https://doi.org/10.1385/1-59259-907-9:215
Download citation
DOI: https://doi.org/10.1385/1-59259-907-9:215
Publisher Name: Humana Press
Print ISBN: 978-1-58829-495-1
Online ISBN: 978-1-59259-907-3
eBook Packages: Springer Protocols