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Journal of Virology, January 2001, p. 251-259, Vol. 75, No. 1
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.1.251-259.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Determination of Coreceptor Usage of Human Immunodeficiency Virus Type 1 from Patient Plasma Samples by Using a Recombinant Phenotypic Assay

Virginie Trouplin,1 Francesca Salvatori,2 Fanny Cappello,1 Veronique Obry,1 Anne Brelot,3 Nikolaus Heveker,3 Marc Alizon,3 Gabriella Scarlatti,2 François Clavel,1 and Fabrizio Mammano1,*

Laboratoire de Recherche Antivirale, INSERM U-82,1 and INSERM U-332 ICGM,3 Paris, France, and Unit of Immunobiology of HIV, DIBIT San Raffaele Scientific Institute, Milan, Italy2

Received 8 August 2000/Accepted 11 October 2000

We developed a recombinant virus technique to determine the coreceptor usage of human immunodeficiency virus type 1 (HIV-1) from plasma samples, the source expected to represent the most actively replicating virus population in infected subjects. This method is not subject to selective bias associated with virus isolation in culture, a step required for conventional tropism determination procedures. The addition of a simple subcloning step allowed semiquantitative evaluation of virus populations with a different coreceptor (CCR5 or CXCR4) usage specificity present in each plasma sample. This procedure detected mixtures of CCR5- and CXCR4-exclusive virus populations as well as dualtropic viral variants, in variable proportions. Sequence analysis of dualtropic clones indicated that changes in the V3 loop are necessary for the use of CXCR4 as a coreceptor, but the overall context of the V1-V3 region is important to preserve the capacity to use CCR5. This convenient technique can greatly assist the study of virus evolution and compartmentalization in infected individuals.

* Corresponding author. Mailing address: Laboratoire de Recherche Antivirale, IMEA/INSERM Hôpital Bichat-Claude Bernard, 46 rue H. Huchard, 75018 Paris, France. Phone: 33-1-4025 6359. Fax: 33-1-4025 6370. E-mail: mammano{at}bichat.inserm.fr.

Journal of Virology, January 2001, p. 251-259, Vol. 75, No. 1
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.1.251-259.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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