Caracterização molecular dos genótipos do rotavírus em pacientes pediátricos imunossuprimidos e não imunossuprimidos / Molecular characterization of rotavirus genotypes in immunosuppressed and non-immunosuppressed pediatric patients

OBJETIVO: Descrever a variabilidade genotípica do rotavírus grupo A (RVA) encontrado em pacientes pediátricos imunocompetentes e imunocomprometidos tratados no Hospital de Clínicas/Universidade Federal do Paraná (HC/UFPR), Curitiba, Paraná. MÉTODOS: Foi realizado um estudo transversal com 1.140 amostras de fezes coletadas, de abril de 2001 a dezembro de 2008, em pacientes ambulatoriais e pacientes hospitalizados com gastroenterite aguda encaminhados ao hospital. As técnicas usadas foram o método da aglutinação do látex e imunoensaio enzimático para diagnóstico de RVA. Foi realizada transcrição reversa, seguida por PCR multiplex semi-nested e sequência de nucleotídeos para caracterização do genótipo. Foram relatados dados de combinações de genótipos, clínicos, epidemiológicos, laboratoriais e sobre a presença de infecções hospitalares. RESULTADOS: Foi analisado um total de 80 amostras de fezes positivas para rotavírus. As associações mais frequentes entre os genótipos G e P foram: G4 P[8] (38,9%), G1 P[8] (30,5%), G9 P[8] (13,9%), G2 P[4] (6.9 %) e G3 P[8] 1,4%). O genótipo prevalente foi G2 P[4] depois da implementação da vacina nos anos de 2006 e 2008. Verificou-se que um total de 62,5% das crianças com idade abaixo de 12 meses estavam infectadas. Destas, 55,6% tinham grave desidratação, e 26,7% precisaram de cuidados intensivos. Encontrou-se uma frequência de 12,5% de infecções hospitalares. Não se observou correlação entre o genótipo e a gravidade da infecção nos pacientes estudados. CONCLUSÃO: As infecções por RVA podem associar-se a manifestações clínicas graves e é crucial a vigilância da variabilidade genotípica desse vírus para monitorizar a emergência de novas cepas e o impacto da imunização nesses pacientes.

OBJECTIVE: To describe the genotypic variability of group A rotavirus (RVA) found in immunosuppressed and non-immunosuppressed pediatric patients treated at the Hospital de Clínicas da Universidade Federal do Paraná (HC-UFPR), Curitiba, Paraná. METHODS: A cross-sectional study was conducted with 1,140 stool samples collected from April, 2001 to December, 2008 in outpatients and hospitalized patients with acute gastroenteritis referred to the hospital. RVA diagnosis was performed through the latex agglutination method and enzyme immunoassay. Reverse transcription followed by multiplex hemi-nested polymerase chain reaction (PCR) and nucleotide sequencing were used for genotype characterization. Genotype combinations, clinical data, epidemiological data, laboratory data, and presence of hospital-acquired infections were reported. RESULTS: A total of 80 rotavirus-positive stool samples were analyzed. The most frequent associations between genotypes G and P were: G4 P[8] (38.9%), G1 P[8] (30.5%), G9 P[8] (13.9%), G2 P[4] (6.9%), and G3 P[8] (1.4%). G2 P[4] was the most prevalent genotype after the vaccine implementation in the years 2006 and 2008. A total of 62.5% of children aged less than 12 months were found to be infected. Of these, 55.6% had severe dehydration and 26.7% needed intensive care. A frequency of 12.5% of nosocomial infections was found. No correlation was observed between genotype and severity of infection in the study patients. CONCLUSION: RVA infections can be associated with severe clinical manifestations, and the surveillance of genotypic variability of this virus is crucial to monitor the emergence of new strains and the impact of the immunization in these patients.

Molecular characterization of rotavirus genotypes in immunosuppressed and non-immunosuppressed pediatric patients.

OBJECTIVE: To describe the genotypic variability of group A rotavirus (RVA) found in immunosuppressed and non-immunosuppressed pediatric patients treated at the Hospital de Clínicas da Universidade Federal do Paraná (HC-UFPR), Curitiba, Paraná. METHODS: A cross-sectional study was conducted with 1,140 stool samples collected from April, 2001 to December, 2008 in outpatients and hospitalized patients with acute gastroenteritis referred to the hospital. RVA diagnosis was performed through the latex agglutination method and enzyme immunoassay. Reverse transcription followed by multiplex hemi-nested polymerase chain reaction (PCR) and nucleotide sequencing were used for genotype characterization. Genotype combinations, clinical, epidemiological, laboratory data, and presence of hospital-acquired infections were reported. RESULTS: A total of 80 rotavirus-positive stool samples were analyzed. The most frequent associations between genotypes G and P were: G4 P[8] (38.9%), G1 P[8] (30.5%), G9 P[8] (13.9%), G2 P[4] (6.9%), and G3 P[8] (1.4%). G2 P[4] was the most prevalent genotype after the vaccine implementation in the years 2006 and 2008. A total of 62,5% of infected children were aged less than 12 months. Of these, 55.6% had severe dehydration and 26.7% needed intensive care. A frequency of 12.5% of nosocomial infections was found. No correlation was observed between genotype and severity of infection in the study patients. CONCLUSION: RVA infections can be associated with severe clinical manifestations, and the surveillance of genotypic variability of this virus is crucial to monitor the emergence of new strains and the impact of the immunization in these patients.

Adenovirus respiratory infection: significant increase in diagnosis using PCR comparing with antigen detection and culture methods / Infecção respiratória aguda por adenovirus: comparação dos métodos de PCR e imunofluorescência indireta para o seu diagnóstico e dados clínicos dos pacientes infectados

Adenovirus (AdV) respiratory infections are usually described as being associated with high mortality rates. Laboratory diagnosis is essential for the establishment of the appropriate therapy, and for guiding the implementation of preventive measures in order to prevent the spread of the infection. Aiming to analyze the sensitivity and specificity of the laboratorial diagnosis methods available, we compared antigen detection by indirect immunofluorescence assay (IF), and a specific nested polymerase chain reaction (PCR), to detect AdV in respiratory samples collected from patients admitted to hospital with acute respiratory disease. Positive samples were inoculated into a cell culture to confirm the results. We analyzed 381 samples from the nasopharyngeal aspirates collected during the year 2008; of these, 2.6 percent tested were positive for adenovirus through IF and 10 percent through PCR; positive isolation was obtained in 40 percent and 26 percent of these cases, respectively. Most infected patients were children under six months of age, and despite of the fact that a significant number of patients required intensive care, the mortality rate was low (5 percent). In conclusion, molecular methods were found to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection; their introduction permitted a significant increase in diagnoses of adenovirus infections.

Infecções respiratórias por Adenovírus (ADV) são geralmente descritas associadas com alta mortalidade. O diagnóstico laboratorial é essencial para o estabelecimento da terapêutica adequada e para orientar a implantação de medidas preventivas evitando a propagação da infecção. Com o objetivo de analisar a sensibilidade e a especificidade dos métodos de avaliação de diagnóstico laboratorial, foi comparada a detecção de antígeno por imunofluorescência indireta (IF) com a reação em cadeia da polimerase específica (PCR) para detectar AdV em amostras respiratórias coletadas de pacientes internados com doença respiratória aguda. As amostras com resultados positivos foram inoculadas em cultura celular. Foram analisadas 381 amostras da secreção nasofaríngea coletadas durante o ano de 2008, das quais 2,6 por cento foram positivas pela IF e 10 por cento pela PCR, isolamento positivo foi obtido em 40 por cento e 26 por cento dos casos positivos pelos testes anteriores, respectivamente. A maioria dos pacientes infectados eram crianças com menos de seis meses de idade, e apesar do fato de que um número significativo de pacientes necessitou de cuidados intensivos, a taxa de mortalidade foi baixa (5 por cento). Em conclusão, os métodos moleculares são úteis para o diagnóstico rápido de infecções por adenovírus com maior sensibilidade do que a detecção do antígeno, a sua introdução na rotina permitiu um aumento significativo no diagnóstico de infecções por adenovírus.

Adenovirus respiratory infection: significant increase in diagnosis using PCR comparing with antigen detection and culture methods.

Adenovirus (AdV) respiratory infections are usually described as being associated with high mortality rates. Laboratory diagnosis is essential for the establishment of the appropriate therapy, and for guiding the implementation of preventive measures in order to prevent the spread of the infection. Aiming to analyze the sensitivity and specificity of the laboratorial diagnosis methods available, we compared antigen detection by indirect immunofluorescence assay (IF), and a specific nested polymerase chain reaction (PCR), to detect AdV in respiratory samples collected from patients admitted to hospital with acute respiratory disease. Positive samples were inoculated into a cell culture to confirm the results. We analyzed 381 samples from the nasopharyngeal aspirates collected during the year 2008; of these, 2.6% tested were positive for adenovirus through IF and 10% through PCR; positive isolation was obtained in 40% and 26% of these cases, respectively. Most infected patients were children under six months of age, and despite of the fact that a significant number of patients required intensive care, the mortality rate was low (5%). In conclusion, molecular methods were found to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection; their introduction permitted a significant increase in diagnoses of adenovirus infections.

Cerebrospinal fluid cytological and biochemical characteristics in the presence of CNS neoplasia.

Central nervous system (CNS) infiltration must be ruled out in patients with known neoplastic diseases and neurological symptoms. It was done a retrospective analysis of 1,948 CSF samples from patients with suspected malignant infiltration in the CNS, in order to evaluate the positivity rate of malignant cells in cerebrospinal fluid (CSF) samples and correlate with cytochemical characteristics. Sixty-two percent of subjects had acute lymphocytic leukemia. Malignant cells were found in 24% of all CSF samples. Subjects with positive malignant cells had predominance of increased levels of CSF total protein (TP), glucose and total cytology (p<0.05). Mean total cell count in this group was 232 (SD 933) cells/mm(3), compared to 9 (SD 93) cells/mm(3) in the group without neoplasic cells (p=0.029). CSF TP specificity was 87% and negative predictive value (NPV) 96%. CSF total cell count specificity 86% and NPV 97%. Although sensitivity and positive predictive value were low. The presence of inflammatory cells and elevated TP found in patients with malignant cells in the CSF can aid in diagnosing CNS neoplasms.

Cerebrospinal fluid cytological and biochemical characteristics in the presence of CNS neoplasia

Central nervous system (CNS) infiltration must be ruled out in patients with known neoplastic diseases and neurological symptoms. It was done a retrospective analysis of 1,948 CSF samples from patients with suspected malignant infiltration in the CNS, in order to evaluate the positivity rate of malignant cells in cerebrospinal fluid (CSF) samples and correlate with cytochemical characteristics. Sixty-two percent of subjects had acute lymphocytic leukemia. Malignant cells were found in 24 percent of all CSF samples. Subjects with positive malignant cells had predominance of increased levels of CSF total protein (TP), glucose and total cytology (p<0.05). Mean total cell count in this group was 232 (SD 933) cells/mm³, compared to 9 (SD 93) cells/mm³ in the group without neoplasic cells (p=0.029). CSF TP specificity was 87 percent and negative predictive value (NPV) 96 percent. CSF total cell count specificity 86 percent and NPV 97 percent. Although sensitivity and positive predictive value were low. The presence of inflammatory cells and elevated TP found in patients with malignant cells in the CSF can aid in diagnosing CNS neoplasms.

A infiltração neoplásica no SNC deve ser afastada em pacientes com neoplasia e sintomas neurológicos. Foi realizada uma análise retrospectiva de 1.948 amostras de LCR de pacientes com suspeita de infiltração neoplásica no SNC. Sessenta e dois por cento dos pacientes eram portadores de leucemia linfocitica aguda. Células neoplásicas foram encontradas em 24 por cento de todas as amostras. Houve níveis aumentados no LCR da proteína total (PT), glicose e de citologia global (p<0.05), no grupo com presença de células neoplásicas. A média da contagem global de células no LCR, neste grupo, foi 232±933 cels/mm³, contra 9±93 cells/mm³ no grupo sem células neoplásicas no LCR (p=0,029). O aumento de PT no LCR apresentou especificidade 87 por cento e valor preditivo negativo (VPN) 96 por cento. A contagem global de células no LCR apresentou especificidade 86 por cento e VPN 97 por cento. Porém sensibilidade e valores preditivos positivos foram baixos. A presença de células inflamatórias e PT no LCR elevada em pacientes com neoplasias pode ser um indicador do envolvimento no SNC.

Acute bacterial meningitis in HIV, patients in southern Brazil: Curitiba, Paraná, Brazil.

Acute communitarian bacterial meningitis and AIDS are prevalent infectious disease in Brazil. The objective of this study was to evaluate the frequency of acute communitarian bacterial meningitis in AIDS patients, the clinical and cerebrospinal fluid (CSF) characteristics. It was reviewed the Health Department data from city of Curitiba, Southern Brazil, from 1996 to 2002. During this period, 32 patients with AIDS fulfilled criteria for acute bacterial meningitis, representing 0.84% of the AIDS cases and 1.85% of the cases of bacterial meningitis. S. pneumoniae was the most frequent bacteria isolated. The number of white blood cells and the percentage of neutrophils were higher and CSF glucose was lower in the group with no HIV co-infection (p 0.12; 0.008; 0.04 respectively). Bacteria not so common causing meningitis can occur among HIV infected patients. The high mortality rate among pneumococcus meningitis patients makes pneumococcus vaccination important.

Acute bacterial meningitis in HIV, pacients in southern Brazil: Curitiba, Paraná, Brazil

Acute communitarian bacterial meningitis and AIDS are prevalent infectious disease in Brazil. The objective of this study was to evaluate the frequency of acute communitarian bacterial meningitis in AIDS patients, the clinical and cerebrospinal fluid (CSF) characteristics. It was reviewed the Health Department data from city of Curitiba, Southern Brazil, from 1996 to 2002. During this period, 32 patients with AIDS fulfilled criteria for acute bacterial meningitis, representing 0.84 percent of the AIDS cases and 1.85 percent of the cases of bacterial meningitis. S. pneumoniae was the most frequent bacteria isolated. The number of white blood cells and the percentage of neutrophils were higher and CSF glucose was lower in the group with no HIV co-infection (p 0.12; 0.008; 0.04 respectively). Bacteria not so common causing meningitis can occur among HIV infected patients. The high mortality rate among pneumococcus meningitis patients makes pneumococcus vaccination important.

A meningite bacteriana aguda comunitária e a AIDS são doenças prevalentes no Brasil. O objetivo desse estudo foi avaliar a freqüência de meningite bacteriana aguda comunitária entre os pacientes com AIDS e as características clínicas e do líquido cefalorraquidiano (LCR). Foram revistos os dados da Secretaria Municipal da Saúde, Curitiba, Paraná, Brasil, nos anos de 1996 a 2002. Nesse período, 32 pacientes com AIDS preencheram os critérios para meningite bacteriana aguda, representando 0,84 por cento dos casos com AIDS e 1,85 por cento dos casos com meningite bacteriana aguda. A bactéria mais freqüentemente isolada foi S. pneumoniae. A celularidade total e a porcentagem de neutrófilos no LCR foi mais elevada e a glicose foi mais baixa no grupo sem co-infecção (p 0,12; 0,008; 0,04 respectivamente). Bactérias menos freqüentes como agentes etiológicos de meningite podem ocorrer. A taxa de mortalidade elevada entre pacientes com meningite por pneumococo torna a vacinação importante.

The Brazilian contribution to the study of neurocysticercosis: Moses and LangeÆs role in cerebrospinal fluid diagnosis

Descrevemos as contribuições pioneiras de Arthur Moses (Instituto Oswaldo Cruz, Rio de Janeiro) em 1911 e de Oswaldo Lange (Faculdade de Medicina da USP, São Paulo) em 1940, para o diagnóstico da neurocisticercose (NC). Moses fez a primeira descrição mundial de um método para o diagnóstico imunológico da NC e Lange fez a descrição da síndrome liquórica da NC.