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Antimicrobial Agents and Chemotherapy, July 1999, p. 1674-1680, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Sequence Diversity of the Reverse Transcriptase of Human Immunodeficiency Virus Type 1 from Untreated Brazilian Individuals

Rodrigo Brindeiro,1 Bart Vanderborght,2,3 Elena Caride,1 Letícia Correa,2 Rejane M. Oravec,4 Oscar Berro,5 Lieven Stuyver,3 and Amilcar Tanuri1,*

Departamento de Genética, Instituto de biologia,1 and Hospital Universitário Clementino Fraga F°,2 Universidade Federal do Rio de Janeiro, and Instituto Noel Nutels,5 Rio de Janeiro, and Hemocentro do Rio Grande do Sul, Rio Grande de Sul,4 Brazil, and Innogenetics, Gent, Belgium3

Received 9 December 1998/Returned for modification 24 February 1999/Accepted 22 April 1999

The presence of human immunodeficiency virus type 1 (HIV-1) bearing mutations resistant to nucleosidic inhibitors of the viral reverse transcriptase (RT) derived from HIV-seropositive asymptomatic and untreated volunteer blood donors was examined. The RT amplicons of 32 specimens were analyzed by using a reverse hybridization line probe assay technique that detects resistance against zidovudine (3'-azido-3'-deoxythymidine [AZT], didanosine (2',3'-dideoxyinosine [ddI], zalcitabine (2',3'-dideoxycytidine [ddC]), and lamivudine {(-)-beta -L-2',3'-dideoxy-3'-thiacytidine [3TC]} at amino acid positions 41, 69, 70, 74, 184, and 215 of the HIV RT. One sample (brp004, subtype B) showed an AZT resistance secondary mutation at position K70R. Fifteen specimens revealed one or more sites of nonreactivity to both wild-type- and mutant-specific probes (dual nonreactivity). Samples were also submitted to RT direct sequencing and phylogenetic analysis. Nine of 32 specimens belonged to non-B subtypes (C, D, F, and F/B or B/F mosaics). Three of these non-B isolates, named brp004, brp063, and brp069, revealed three other relevant AZT resistance mutations---a T215F mutation and two M41L mutations, respectively---hidden by the nonreactivity to line probe assay strips on the respective codon regions. The isolate brp004 also carried a D67N AZT resistance mutation revealed by direct sequencing. No nonnucleosidic RT inhibitor-resistant mutation was found. The analysis revealed a frequency of 2.26 × 10-4 mutations per nucleotide for independent samples related to RT resistance. These findings emphasize the magnitude of naturally occurring reservoirs of drug-resistant virus among untreated HIV-1-positive individuals in Brazil.

* Corresponding author. Mailing address: Lab. Virologia Molecular, Depto. de Genética, sala A2-121, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro (UFRJ), av. Brig. Trompowski s/n, CEP 21941-590 Rio de Janeiro, RJ, Brazil. Phone: 55 021 280-8043 R42. Fax: 55 021 205-2671. E-mail: lavimoan{at}hotmail.com.

Antimicrobial Agents and Chemotherapy, July 1999, p. 1674-1680, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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