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Journal of Clinical Microbiology, June 2001, p. 2110-2114, Vol. 39, No. 6
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.6.2110-2114.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Development of an env gp41-Based Heteroduplex Mobility Assay for Rapid Human Immunodeficiency Virus Type 1 Subtyping

S. M. Agwale,^1,2 K. E. Robbins,¹ L. Odama,³ A. Saekhou,¹ C. Zeh,¹ A. Edubio,^3, O. M. Njoku,⁴ N. Sani-Gwarzo,⁵ M. F. Gboun,⁵ F. Gao,⁶ M. Reitz,² D. Hone,² T. M. Folks,¹ D. Pieniazek,¹ C. Wambebe,³ and M. L. Kalish^1,*

Centers for Disease Control and Prevention, Atlanta, Georgia¹; Division of Vaccine Research, Institute of Human Virology, Baltimore, Maryland²; National Institute for Pharmaceutical Research and Development³ and Federal Ministry of Health,⁵ Abuja, Nigeria; Robert-Koch Institut, Berlin, Germany⁴; and University of Alabama at Birmingham, Birmingham, Alabama⁶

Received 26 October 2000/Returned for modification 4 January 2001/Accepted 7 March 2001

The gp120 region of the human immunodeficiency virus type 1 (HIV-1) envelope (env) gene exhibits a high level of genetic heterogeneity across the group M subtypes. The heteroduplex mobility assay (HMA) has successfully been used to assign subtype classifications, but C2V5 primers often fail to amplify African strains. We developed an env gp41-based HMA for which the target sequence is amplified with highly conserved gp41 primers, known to efficiently amplify nucleic acids from HIV-1 group M, N, and O viruses. By using gp41 from a new panel of reference strains, the subtype assignments made by our modified HMA were concordant with those obtained by sequencing and phylogenetic analysis of 34 field strains from 10 countries representing subtypes A to G. Testing of field strains from Nigeria further demonstrated the utility of this modified assay. Of 28 samples, all could be amplified with gp41 primers but only 17 (60.7%) could be amplified with the standard C2V5 primers. Therefore, gp41-based HMA can be a useful tool for the rapid monitoring of prevalent subtypes in countries with divergent strains of circulating HIV-1.

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^* Corresponding author. Mailing address: Centers for Disease Control and Prevention, Mail Stop G19, 1600 Clifton Rd., Atlanta, GA 30333. Phone: (404) 639-3957. Fax: (404) 639-2919. E-mail: Mkalish{at}cdc.gov.

Present address: Robert Koch Institute, Berlin, Germany.

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Journal of Clinical Microbiology, June 2001, p. 2110-2114, Vol. 39, No. 6
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.6.2110-2114.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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