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J Virol Methods ; 128(1-2): 47-53, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15871907

RESUMO

Genotypic assays are used often to guide clinicians in decisions concerning the treatment of patients. An optimized sequence-based genotypic assay was used to determine the whole protease and reverse transcriptase (RT) gene, including the gag cleavage site region and RNase H region. Since non-B subtypes are increasing in countries where subtype B was the most prevalent subtype, and treatment becomes more available in developing countries where the epidemic is characterized by a high prevalence of non-B subtypes, it was important that the genotypic test was evaluated using a panel of different subtypes. Amplification was successful for different subtypes: A, B, C, D, F, G, H, J, CRF01_AE, CRF02_AG, CRF11_cpx, CRF13_cpx and an uncharacterized recombinant sample. The detection limit of the PCR was 1000 copies/ml, except for 1 subtype C sample (PL3) and 1 CRF02_AG sample (PL8). The detection limit for these samples was 5000 copies/ml. A sequence could be obtained in both directions for most of the samples.


Assuntos
Infecções por HIV/virologia , Protease de HIV/classificação , Transcriptase Reversa do HIV/classificação , HIV-1/classificação , HIV-1/genética , Reação em Cadeia da Polimerase/métodos , Primers do DNA , DNA Complementar/metabolismo , Farmacorresistência Viral/genética , Produtos do Gene gag/química , Produtos do Gene gag/metabolismo , Genótipo , Protease de HIV/genética , Transcriptase Reversa do HIV/genética , HIV-1/efeitos dos fármacos , HIV-1/enzimologia , Humanos , RNA Viral/isolamento & purificação , Ribonuclease H/genética
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