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Journal of Virology, January 2000, p. 363-370, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Simplified Strategy for Detection of Recombinant Human Immunodeficiency Virus Type 1 Group M Isolates by gag/env Heteroduplex Mobility Assay

Leo Heyndrickx,1 Wouter Janssens,1,2,* Léopold Zekeng,3 Rosemary Musonda,4 Séverin Anagonou,5 Gert Van der Auwera,1 Sandra Coppens,1 Katleen Vereecken,1 Ko De Witte,1 Rian Van Rampelbergh,1 Maina Kahindo,6 Linda Morison,7 Francine E. McCutchan,8 Jean K. Carr,8 Jan Albert,9 Max Essex,10 Jaap Goudsmit,11 Birgitta Asjö,12 Mika Salminen,13 Anne Buvé,1 Study Group on Heterogeneity of HIV Epidemics in African Cities,dagger and Guido van der Groen1

Department of Microbiology, Institute of Tropical Medicine, Antwerp,1 and The Flanders Interuniversity Institute for Biotechnology (VIB), Zwijnaarde,2 Belgium; Laboratoire de Santé Hygiène Mobile, Ministère de la Santé, Yaoundé, Cameroon3; Tropical Diseases Research Centre, Ndola, Zambia4; Programme National de Lutte contre le SIDA, Cotonou, Bénin5; National AIDS/STD Control Programme, Nairobi, Kenya6; London School of Hygiene & Tropical Medicine, Department of Infections and Tropical Diseases, London, United Kingdom7; Henry M. Jackson Foundation, Rockville, Maryland8; Swedish Institute for Infectious Disease Control, Karolinska Institute, Stockholm, Sweden9; Harvard Institute and the Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts10; Department of Human Retrovirology, Amsterdam, The Netherlands11; National Centre for Research in Virology, University of Bergen, Bergen, Norway12; and Department of Chronic Viral Infections, National Public Health Institute, Helsinki, Finland13

Received 10 May 1999/Accepted 22 September 1999

We developed a heteroduplex mobility assay in the gag gene (gag HMA) for the identification of group M subtypes A to H. The assay covers the region coding for amino acid 132 of p24 to amino acid 20 of p7 (according to human immunodeficiency virus type 1 [HIV-1] ELI, 460 bp). The gag HMA was compared with sequencing and phylogenetic analysis of an evaluation panel of 79 HIV-1 group M isolates isolated from infected individuals from different geographic regions. Application of gag HMA in combination with env HMA on 252 HIV-1- positive plasma samples from Bénin, Cameroon, Kenya, and Zambia revealed a high prevalence of a variety of intersubtype recombinants in Yaoundé, Cameroon (53.8%); Kisumu, Kenya (26.8%); and Cotonou, Bénin (41%); no recombinants were identified among the samples from Ndola, Zambia. The AGIbNG circulating recombinant form, as determined by gag HMA, was found to be the most common intersubtype recombinant in Yaoundé (39.4%) and Cotonou (38.5%). Using a one-tube reverse transcriptase PCR protocol, this gag HMA in combination with env HMA is a useful tool for rapidly monitoring the prevalence of the various genetic subtypes as well as of recombinants of HIV-1. Moreover, this technology can easily be applied in laboratories in developing countries.

* Corresponding author. Mailing address: Department of Microbiology, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium. Phone: 32-3-247-63-28. Fax: 32-3-247-63-33. E-mail: wjanssens{at}itg.be.

dagger Members of the Study Group on Heterogeneity of HIV Epidemics in African Cities include A. Buvé (coordinator), M. Laga, E. Van Dyck, W. Janssens, and L. Heyndrickx (Institute of Tropical Medicine, Antwerp, Belgium); M. Caraël (UNAIDS); S. Anagonou (Programme National de Lutte contre le SIDA, Bénin); M. Laourou (Institut National de Statistiques et d'Analyses Economiques, Bénin); L. Kanhonou (Centre de Recherche en Reproduction Humaine et en Démographie, Bénin); L. Zekeng (Laboratoire de Santé Hygiène Mobile, Cameroon); E. Akam and M. de Loenzien (Institut de Formation et de Recherche en Démographiques, Cameroon); S.-C. Abega (Université Catholique d'Afrique Centrale, Cameroon); M. Kahindo (formerly National AIDS/STD Control Programme, Kenya); J. Chege and N. Rutenberg (The Population Council, Nairobi); V. Kimani (Department of Community Health, University of Nairobi); R. Musonda, T. Sukwa, and F. Kaona (Tropical Diseases Research Centre, Zambia); B. Auvert and E. Lagarde (INSERM U88, Paris, France); N. J. Robinson (formerly INSERM U88, Paris, France); B. Ferry and N. Lydié (Centre Français sur la Population et le Développement, Paris, France); and R. Hayes, L. Morison, H. Weiss, and J. Glynn (London School of Hygiene and Tropical Medicine, London, United Kingdom).

Journal of Virology, January 2000, p. 363-370, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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