ABSTRACT
Objective: To evaluate genotyping and subtyping in antiretroviral (ARV) naïve and experienced children, as well as drug resistance profiles through genotyping in these children. Methods: This retrospective study assessed ARV-naïve HIV children and HIV children failing highly active antiretroviral treatment (HAART) followed up at Santa Casa de São Paulo. Genotypingwas performed using purified polymerase chain reaction (PCR) products from retrotranscribed RNA using Kit Viroseq HIV-1 Genotyping System 2.0 or nested PCR in-house. Sequencing was performed using automatic equipment (ABI 3100). ARV resistance mutations were analyzed in the Stanford HIV Drug Resistance Database and subtypingwas performed at the National Center for Biotechnology Information (NCBI), using SimPlot analysis, together with phylogenetic analysis. Results: No primary ARV resistance mutation was detected in the 24 ARV-naïve children, although there were mutations that may contribute to resistance to nucleoside analogue reverse transcriptase inhibitors (NRTI) (12.5%) and to protease inhibitors (PI) (95.8%). For the 23 children failing HAART, we found ARV resistance mutations to NRTI in 95.6% and to non-nucleoside analogue reverse transcriptase inhibitors (NNRTI) in 60.8%. For PI, we found ARV resistance mutations in 95.7%, 47.8% of which had only polymorfisms. In the subtyping analyses, 78.3% of the sequences clustered in HIV-1 subtype B, 4.3% in C, 13% in F and 4.4% in recombinant forms. Conclusion: Our results show low rates of primary resistance in ARV-naïve children and high rates of resistance in children failing ARV treatment, which is compatible with ARV use in these patients.
Objetivo: Avaliar a genotipagem e subtipagem em crianças experimentadas e virgens de tratamento, assimcomoperfis de resistência a medicamentos através da genotipagem nessas crianças. Métodos: Estudo retrospectivo de crianças HIV positivas virgens de tratamento e HIV positivas que não responderam ao tratamento pela terapia antirretroviral altamente ativa (HAART), acompanhadas na Santa Casa de São Paulo (SP). A genotipagem foi realizada com produtos purificados de reação em cadeia da polimerase (PCR) de RNA retrotranscrito, utilizando-se o kit comercial Viroseq HIV-1 Genotyping System 2.0 ou a técnica de nested PCR in-house. O sequenciamento foi realizado com equipamento automático (ABI 3100). As mutações de resistência antirretroviral (ARV) foram analisadas no Stanford HIV Drug Resistance Database e a subtipagem realizada no U.S. National Center for Biotechnology Information (NCBI), utilizando-se o programa de análises SimPlot, juntamente com a análise filogenética. Resultados: Não foi detectada nenhuma mutação de resistência primária ARV nas 24 crianças virgens de tratamento, embora tenham ocorrido mutações que podem contribuir para a resistência aos inibidores da transcriptase reversa análogos de nucleosídeos (ITRN) (12,5%) e aos inibidores da protease (IP) (95,8%). Para as 23 crianças que não responderam à HAART, foram encontradas mutações de resistência ARV aos ITR Nem 95,6% e aos inibidores da transcriptase reversa não-análogos de nucleosídeos (ITRNN) em 60,8%. Para os IP, foram observadas mutações de resistência ARV em 95,7%, 47,8% das quais apresentavam apenas polimorfismos. Nas análises de subtipagem,78,3%das sequências agruparam-se no subtipo B do HIV-1, 4,3% no C, 13% no F e 4,4% em formas recombinantes. Conclusões: Nossos resultados mostrambaixas taxas de resistência primária em crianças virgens de tratamento e altas taxas de resistência emcrianças que não responderamao tratamento ARV, o que é compatível com o uso ARV nesses pacientes.
ABSTRACT
OBJETIVO: Avaliar a genotipagem e subtipagem em crianças experimentadas e virgens de tratamento, assim como perfis de resistência a medicamentos através da genotipagem nessas crianças. MÉTODOS: Estudo retrospectivo de crianças HIV positivas virgens de tratamento e HIV positivas que não responderam ao tratamento pela terapia antirretroviral altamente ativa (HAART), acompanhadas na Santa Casa de São Paulo (SP). A genotipagem foi realizada com produtos purificados de reação em cadeia da polimerase (PCR) de RNA retrotranscrito, utilizando-se o kit comercial Viroseq HIV-1 Genotyping System 2.0 ou a técnica de nested PCR in-house. O sequenciamento foi realizado com equipamento automático (ABI 3100). As mutações de resistência antirretroviral (ARV) foram analisadas no Stanford HIV Drug Resistance Database e a subtipagem realizada no U.S. National Center for Biotechnology Information (NCBI), utilizando-se o programa de análises SimPlot, juntamente com a análise filogenética. RESULTADOS: Não foi detectada nenhuma mutação de resistência primária ARV nas 24 crianças virgens de tratamento, embora tenham ocorrido mutações que podem contribuir para a resistência aos inibidores da transcriptase reversa análogos de nucleosídeos (ITRN) (12,5%) e aos inibidores da protease (IP) (95,8%). Para as 23 crianças que não responderam à HAART, foram encontradas mutações de resistência ARV aos ITRN em 95,6% e aos inibidores da transcriptase reversa não-análogos de nucleosídeos (ITRNN) em 60,8%. Para os IP, foram observadas mutações de resistência ARV em 95,7%, 47,8% das quais apresentavam apenas polimorfismos. Nas análises de subtipagem, 78,3% das sequências agruparam-se no subtipo B do HIV-1, 4,3% no C, 13% no F e 4,4% em formas recombinantes. CONCLUSÕES: Nossos resultados mostram baixas taxas de resistência primária em crianças virgens de tratamento e altas taxas de resistência...
OBJECTIVE: To evaluate genotyping and subtyping in antiretroviral (ARV) naïve and experienced children, as well as drug resistance profiles through genotyping in these children. METHODS: This retrospective study assessed ARV-naïve HIV children and HIV children failing highly active antiretroviral treatment (HAART) followed up at Santa Casa de São Paulo. Genotyping was performed using purified polymerase chain reaction (PCR) products from retrotranscribed RNA using Kit Viroseq HIV-1 Genotyping System 2.0 or nested PCR in-house. Sequencing was performed using automatic equipment (ABI 3100). ARV resistance mutations were analyzed in the Stanford HIV Drug Resistance Database and subtyping was performed at the National Center for Biotechnology Information (NCBI), using SimPlot analysis, together with phylogenetic analysis. RESULTS: No primary ARV resistance mutation was detected in the 24 ARV-naïve children, although there were mutations that may contribute to resistance to nucleoside analogue reverse transcriptase inhibitors (NRTI) (12.5%) and to protease inhibitors (PI) (95.8%). For the 23 children failing HAART, we found ARV resistance mutations to NRTI in 95.6% and to non-nucleoside analogue reverse transcriptase inhibitors (NNRTI) in 60.8%. For PI, we found ARV resistance mutations in 95.7%, 47.8% of which had only polymorfisms. In the subtyping analyses, 78.3 percent of the sequences clustered in HIV-1 subtype B, 4.3% in C, 13% in F and 4.4% in recombinant forms. CONCLUSION: Our results show low rates of primary resistance in ARV-naïve children and high rates of resistance in children failing ARV treatment, which is compatible with ARV use in these patients.
Subject(s)
Adolescent , Child , Child, Preschool , Humans , Infant , Infant, Newborn , Antiretroviral Therapy, Highly Active , Drug Resistance, Viral/genetics , HIV Infections/virology , HIV-1 , Mutation , Genotype , HIV Infections/drug therapy , HIV Reverse Transcriptase/genetics , HIV-1 , Phylogeny , Polymerase Chain Reaction , Prevalence , Retrospective StudiesABSTRACT
Streptococcus pyogenes meningitis (SPM) occurs sporadically, even with the increase of invasive streptococcal disease observed in the past years. We reported two cases of SPM in infants to alert pediatricians for the possibility of this agent as a cause of meningitis in previously healthy children.
Subject(s)
Child, Preschool , Humans , Infant , Male , Meningitis, Bacterial/microbiology , Streptococcal Infections/microbiology , Streptococcus pyogenes/isolation & purification , Meningitis, Bacterial/diagnosis , Meningitis, Bacterial/drug therapy , Penicillins/therapeutic use , Streptococcal Infections/diagnosis , Streptococcal Infections/drug therapyABSTRACT
The aim of this study was to identify the risk factors for nosocomial bloodstream infections by multidrug resistant Gram-negative bacilli. From November 2001 to December 2003, in the Pediatric Department of the Santa Casa de São Paulo, a retrospective case-control study was developed concerning patients who had nosocomial bloodstream infection caused by Gram-negative bacilli. Patients with multidrug resistant infections were designated as case patients, and control patients were those with an infection that did not meet the criteria for multidrug resistance. Previous use of central venous catheter and previous use of vancomycin plus third generation cephalosporins were associated to a higher chance of infections by multidrug resistant Gram-negative bacilli (Odds ratio - 5.8 and 5.2, respectively). Regarding sensitivity of the isolated agents, 47.8 percent were multidrug resistant, 54.2 percent were Klebsiella spp. ESBL producers and 36.4 percent were imipenem resistant Pseudomonas aeruginosa. The lethality rate was 36.9 percent in the studied cases and this rate was significantly higher in the group of patients with multidrug resistant infections (p=0.013). Risk factor identification as well as the knowledge of the susceptibility of the nosocomial infectious agents gave us the possibility to perform preventive and control strategies to reduce the costs and mortality related to these infections.