Anti-HIV drugs have recently become available for the treatment of children infected with HIV in Vietnam; however, the genetic background of HIV-1 drug resistance in antiretroviral-naive children has yet to be studied. Of the 104 HIV-1 CRF01-AE subtype strains that were previously isolated from antiretroviral-naive children from the provinces of southern Vietnam and hospitalized in Children Hospital 1 in Ho Chi Minh City from 2004 to 2005, 79 strains were used for amplification and sequence analyses of the protease and reverse transcriptase (RT) genes. Minor mutations were found in the protease gene, including L10I, I13V, G16E, M36I, D60E, I62V, I64V, L63P, H69K, V82I, and I93L. Of these mutations, M36I and H69K were detected in all of the strains that were studied. However, all of the amino acid changes in the protease gene were considered to be polymorphisms. In the RT gene, three major mutations were detected in six strains: the V75M mutation in one strain, the Y181C mutation in two strains, and the M184I mutation in three strains. The prevalence of primary or transmitted HIV drug resistance to all of the drugs and drug classes that were evaluated in this study was 7.6%. These findings provide a useful background for antiretroviral therapy in Vietnam and contribute reference data for the surveillance of HIV drug resistance around the world. This study suggests that the prevalence of HIVDR in Vietnam may have recently increased. The monitoring of HIV drug resistance in Vietnam is necessary.