RESUMO
The assessment of potential "breakthrough infections" in HIV vaccine trials requires knowledge of viral load in unvaccinated persons. Therefore, HIV-1 RNA was quantitated in plasma from Thai adults with subtype E infections. RNA was detectable (> or =500 copies/ml) in 93% of 255 specimens, with a mean (standard deviation) value of 4.09 (0.88) log copies/ml. The concentration of RNA was directly related to the presence of AIDS-defining illnesses, inversely related to CD4 count, and independent of gender after adjustment for CD4 count.
Assuntos
Infecções por HIV/virologia , HIV-1/classificação , HIV-1/isolamento & purificação , Carga Viral , Vacinas contra a AIDS/imunologia , Síndrome da Imunodeficiência Adquirida/imunologia , Síndrome da Imunodeficiência Adquirida/virologia , Contagem de Linfócito CD4 , Feminino , Infecções por HIV/imunologia , HIV-1/genética , HIV-1/imunologia , Humanos , Masculino , RNA Viral/análise , RNA Viral/genética , Caracteres Sexuais , Tailândia , VacinaçãoRESUMO
Human immunodeficiency virus type 1 (HIV) RNA load was measured in paired samples of peripheral blood plasma and nasopharyngeal (NP) washes from 97 Thai subjects infected with subtype E or B. HIV RNA was quantifiable in 93% of peripheral blood plasma samples tested and was inversely correlated (rho =-0.524; p < 0.001) with CD4 absolute count. HIV RNA was quantifiable in 29% of NP samples tested, and the median value was less than that of plasma viral load. HIV RNA load in NP samples was correlated (rho = 0.388; p < 0.001) with viral load in peripheral blood. HIV RNA was not detected in NP washes from subjects with undetectable plasma viral load. Virus isolation attempts on two NP samples were negative. The results do not support local HIV production in the nasopharynx, but extend current knowledge of HIV shedding to include the NP compartment.
Assuntos
Infecções por HIV/virologia , HIV-1/isolamento & purificação , Nasofaringe/virologia , RNA Viral/análise , Contagem de Linfócito CD4 , Feminino , HIV-1/genética , HIV-1/fisiologia , Humanos , Masculino , Líquido da Lavagem Nasal/virologia , RNA Viral/sangue , Tailândia , Carga ViralRESUMO
Dried blood spot (DBS) specimens were assessed as an alternative to plasma for human immunodeficiency virus type 1 (HIV-1) serotyping by V3 loop peptide enzyme immunoassay. Nested PCR capable of distinguishing HIV-1 subtypes B and E was used as the reference standard. Ninety-two percent of DBS samples were typeable as either HIV-1 subtype B or E. Serotype results with DBS and plasma were identical for 254 of 257 specimens. A simple DBS collection method provides a convenient alternative for conducting HIV-1 serotype surveillance while retaining sensitivity and specificity.
Assuntos
Síndrome da Imunodeficiência Adquirida/sangue , HIV-1/classificação , HIV-1/isolamento & purificação , Síndrome da Imunodeficiência Adquirida/virologia , Sequência de Aminoácidos , Coleta de Amostras Sanguíneas/métodos , Anticorpos Anti-HIV/sangue , Antígenos HIV/química , Antígenos HIV/imunologia , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Sorotipagem/métodos , TailândiaRESUMO
The aim of this study was to compare the performance of differential polymerase chain reaction (PCR) typing and peptide enzyme-linked immunosorbent assay (V3-EIA) for human immunodeficiency virus type 1 (HIV-1) subtyping in Thailand using heteroduplex mobility assay (HMA) as the reference standard. Paired peripheral blood mononuclear cells (PBMC) and sera were collected from 38 HIV-1 seropositive persons in Thailand. HMA was done by standard methods; differential PCR employs primer pairs that differentially amplify either subtype E or B. V3-EIA used peptides specific for subtypes E or B. Thirty-two cases (84%) were found by HMA to be infected with subtype E: and six with (16%) subtype B. The results obtained with differential PCR were 100% concordant with those of HMA; V3 EIA correctly predicted the subtype in 95% (36 of 38). Six samples that molecularly subtyped as E were repeatedly dual reactive by screening V3-EIA, but these resolved to subtype E using an antigen-limiting EIA. Two samples were serologically nontypeable because of overall low levels of V3 antibody. Using HMA as the standard, differential PCR was shown to subtype HIV-1 reliably from patient PBMC samples. V3-EIA correctly predicted HIV-1 subtype in most (95%) of our cases. Because of the less rigorous sampling requirements, specimen processing, and logistical and technical requirements of serotyping compared with molecular techniques, it appears to be practical for screening purposes in a field environment. Samples that cannot be definitively subtyped serologically should undergo differential PCR and antigen-limiting V3 EIA. These approaches to HIV-1 subtyping should be used in complementary fashion in Thailand, where subtypes B and E are currently known to cocirculate.