Long-term effect of 10-valent pneumococcal conjugate vaccine on nasopharyngeal carriage of Streptococcus pneumoniae in children in Brazil.

Brazil introduced the 10-valent pneumococcal vaccine (PCV10) to the routine national immunization program (NIP) in March 2010. In 2017, we investigated the effects of PCV10 on nasopharyngeal carriage of vaccine-types (VT) and non-vaccine-types (NVT) of Streptococcus pneumoniae (Spn) among children living in São Paulo city. We also compared the prevalence of VT and NVT with previous carriage surveys performed in 2010 (baseline) and 2013. METHOD: The carriage survey was conducted among 531 children, aged 12 months to <24 months, recruited from public Primary Health Units during the immunization campaign, using previous surveys methodology, except for qPCR, which was performed in the 2017 survey only. RESULTS: No statistical difference was found in the prevalence of Spn either by culture (59.7%) or by qPCR (61.2%). Spn carriage increased from 40.3% (baseline) to 59.7% (2017 survey) (p < 0.001). Colonization by VT isolates significantly decreased by 90.9% (19.8-1.8%) and 95.5% (19.8-0.9%) in the 2013 and 2017 surveys, respectively, compared to that at baseline. NVT isolates increased significantly by 128% (19.6-44.8%) and 185% (19.6-55.9%) in the respective post-PCV10 surveys, most led to high prevalence of serotypes 6C (27%), 15B (9.8%), 19A (9.2%), 15A (6.0%), and 16F (5.7%). In 2017, reduction in serotype 6A (4.2-0.6%, p < 0.001) and increase in serotype 19A (1.8-6.0%, p = 0.001) were found; serotype 3 isolate was not detected in the present survey. We identified the emergence of 19A isolates CC320, associated with high penicillin (MIC ≥ 2.0 mg/L) and cefotaxime (MIC ≥ 1.0 mg/L) values. CONCLUSION: After 7 years of PCV10 introduction in the NIP, colonization by VT among toddlers decreased substantially to a residual level, along with substantial serotype replacement by novel serotypes not present in any current conjugated pneumococcal vaccine and serotype 19A. The present findings can assist policy decisions in Brazil.

Long-term effect of 10-valent pneumococcal conjugate vaccine on nasopharyngeal carriage of Streptococcus pneumoniae in children in Brazil

Brazil introduced the 10-valent pneumococcal vaccine (PCV10) to the routine national immunization program (NIP) in March 2010. In 2017, we investigated the effects of PCV10 on nasopharyngeal carriage of vaccine-types (VT) and non-vaccine-types (NVT) of Streptococcus pneumoniae (Spn) among children living in São Paulo city. We also compared the prevalence of VT and NVT with previous carriage surveys performed in 2010 (baseline) and 2013. Method: The carriage survey was conducted among 531 children, aged 12 months to <24 months, recruited from public Primary Health Units during the immunization campaign, using previous surveys methodology, except for qPCR, which was performed in the 2017 survey only. Results: No statistical difference was found in the prevalence of Spn either by culture (59.7%) or by qPCR (61.2%). Spn carriage increased from 40.3% (baseline) to 59.7% (2017 survey) (p < 0.001). Colonization by VT isolates significantly decreased by 90.9% (19.8­1.8%) and 95.5% (19.8­0.9%) in the 2013 and 2017 surveys, respectively, compared to that at baseline. NVT isolates increased significantly by 128% (19.6­44.8%) and 185% (19.6­55.9%) in the respective post-PCV10 surveys, most led to high prevalence of serotypes 6C (27%), 15B (9.8%), 19A (9.2%), 15A (6.0%), and 16F (5.7%). In 2017, reduction in serotype 6A (4.2­0.6%, p < 0.001) and increase in serotype 19A (1.8­6.0%, p = 0.001) were found; serotype 3 isolate was not detected in the present survey. We identified the emergence of 19A isolates CC320, associated with high penicillin (MIC 2.0 mg/L) and cefotaxime (MIC 1.0 mg/L) values. Conclusion: After 7 years of PCV10 introduction in the NIP, colonization by VT among toddlers decreased substantially to a residual level, along with substantial serotype replacement by novel serotypes not present in any current conjugated pneumococcal vaccine and serotype 19A. The present findings can assist policy decisions in Brazil

Incorporation of real-time PCR into routine public health surveillance of culture negative bacterial meningitis in São Paulo, Brazil.

Real-time (RT)-PCR increases diagnostic yield for bacterial meningitis and is ideal for incorporation into routine surveillance in a developing country. We validated a multiplex RT-PCR assay for Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae in Brazil. Risk factors for being culture-negative, RT-PCR positive were determined. The sensitivity of RT-PCR in cerebrospinal fluid (CSF) was 100% (95% confidence limits, 96.0%-100%) for N. meningitidis, 97.8% (85.5%-99.9%) for S. pneumoniae, and 66.7% (9.4%-99.2%) for H. influenzae. Specificity ranged from 98.9% to 100%. Addition of RT-PCR to routine microbiologic methods increased the yield for detection of S. pneumoniae, N. meningitidis, and H. influenzae cases by 52%, 85%, and 20%, respectively. The main risk factor for being culture negative and RT-PCR positive was presence of antibiotic in CSF (odds ratio 12.2, 95% CI 5.9-25.0). RT-PCR using CSF was highly sensitive and specific and substantially added to measures of meningitis disease burden when incorporated into routine public health surveillance in Brazil.

Incorporation of real-time PCR into routine public health Surveillance of culture negative bacterial meningitis in Sao Paulo, Brazil

Real-time (RT)-PCR increases diagnostic yield for bacterial meningitis and is ideal for incorporation into routine surveillance in a developing country. We validated a multiplex RT-PCR assay for Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae in Brazil. Risk factors for being culture-negative, RT-PCR positive were determined. The sensitivity of RT-PCR in cerebrospinal fluid (CSF) was 100% (95% confidence limits, 96.0%–100%) for N. meningitidis, 97.8% (85.5%–99.9%) for S. pneumoniae, and 66.7% (9.4%–99.2%) for H. influenzae. Specificity ranged from 98.9% to 100%. Addition of RT-PCR to routine microbiologic methods increased the yield for detection of S. pneumoniae, N. meningitidis, and H. influenzae cases by52%, 85%, and 20%, respectively. The main risk factor for being culture negative and RT-PCR positive was presence of antibiotic in CSF (odds ratio 12.2, 95% CI 5.9-25.0). RT-PCR using CSF was highly sensitive and specific and substantially added to measures of meningitis disease burden when incorporated into routine public health surveillance in Brazil

Limitações no uso da técnica de contraimunoeletroforese (CIE) para o diagnóstico das meningites causadas por Haemophilus influenzae tipo b / Limitations in the use of counterimmunoelectrophoresis (CIE) for diagnosis of Haemophilus influenza type b meningitis

A contraimunoeletroforese (CIE) é uma técnica amplamente utilizada no Brasil para o diagnóstico laboratorial indireto de meningites causadas por Neisseria meningitidis (Men) dos sorogrupos A, B e C e Haemophilus influenzae (Hi) tipo b, desde a década de 1970. A introdução da técnica de PCR em tempo real (RT-PCR) na rotina diagnóstica das meningites causadas por Men, Hi e Streptococcus pneumoniae (Spn), no Instituto Adolfo Lutz, levou à identificação de resultados discrepantes entre as duas metodologias. O objetivo deste trabalho foi investigar 46 amostras com resultados de CIE positivos para Hib. Deste total, 26 amostras (57%) tiveram resultados caracterizados como falsos positivos para Hib, pois nenhuma delas foi positiva para este agente por RT-PCR e teste de látex. Destas, 21 (46%) foram positivas para Spn por RT-PCR e látex e 5 (11%) foram negativas tanto para Hib ou Spn por ambas as técnicas. Estes dados evidenciaram a alta porcentagem de resultados falsos positivos para o componente Hib obtidos pela técnica de CIE. Nós recomendamos o uso do látex ou RT-PCR e não a CIE para a detecção de Hib ou, então, o uso de um segundo teste para confirmar casos de Hib positivos por CIE.

Limitações no uso da técnica de contraimunoeletroforese (CIE) para o diagnóstico das meningites causadas por Haemophilus influenzae tipo b / limitations in the use of couterimmunoelectrophoresis (CIE) for diagnosis of Haemophilus influenzae type b meningitis

A contraimunoeletroforese (CIE) é uma técnica amplamente utilizada noBrasil para o diagnóstico laboratorial indireto de meningites causadas porNeisseria meningitidis (Men) dos sorogrupos A, B e C e Haemophilusinfluenzae (Hi) tipo b, desde a década de 1970. A introdução da técnica dePCR em tempo real (RT-PCR) na rotina diagnóstica das meningitescausadas por Men, Hi e Streptococcus pneumoniae (Spn), no InstitutoAdolfo Lutz, levou à identificação de resultados discrepantes entre as duasmetodologias. O objetivo deste trabalho foi investigar 46 amostras comresultados de CIE positivos para Hib. Deste total, 26 amostras (57%)tiveram resultados caracterizados como falsos positivos para Hib, poisnenhuma delas foi positiva para este agente por RT-PCR e teste de látex.Destas, 21 (46%) foram positivas para Spn por RT-PCR e látex e 5 (11%)foram negativas tanto para Hib ou Spn por ambas as técnicas. Estes dadosevidenciaram a alta porcentagem de resultados falsos positivos para ocomponente Hib obtidos pela técnica de CIE. Nós recomendamos o uso dolátex ou RT-PCR e não a CIE para a detecção de Hib ou, então, o uso de umsegundo teste para confirmar casos de Hib positivos por CIE

Síndrome de Chediak-Higashi: relato de caso e revisäo de literatura / Chediak-Higashi syndrome: case report and literature review

Objetivo: Apresentar o caso de paciente parda portadora de síndrome de Chediak-Higashi (SCH). Método: Além dos dados de história clínica e de exame físico a paciente foi submetida a investigação laboratorial para imunodeficiência. Resultados: As principais manifestações clínicas da paciente eram albinismo parcial óculo-cutâneo e infecções de repetição. Os exames laboratoriais revelaram a presença de inclusões intracitoplasmáticas gigantes em leucócitos do sangue periférico e em células precursoras na medula óssea. A avaliação funcional dos leucócitos do sangue periférico foi normal. Quatro meses após o diagnóstico, a criança desenvolveu a fase linfoproliferativa evoluindo para óbito. Conclusões: São discutidas as principais alterações laboratoriais que caracterizam a SCH, tece-se comentários sobre os possíveis fatores etiológicos, bem como os avanços no seu tratamento.

Delayed phagocytosis and bacterial killing in Chediak-Higashi syndrome neutrophils detected by a fluorochrome assay: ultrastructural aspects

The few studies already published about phagocyte functions in Chediak-Higashi syndrome (CHS) has stated that neutrophils present slow rate of bacterial killing but normally ingest microorganisms. In the present study, both phagocytosis and killing of Staphylococcus aureus were verified to be in neutrophils from two patients with CHS when these functions were simultaneously evaluated by a fluorochrome phagocytosis assay. Electron microscopic examination showed morphologic differences among neutophils from CHS patients and normal neutrophils regarding the cytoplasmic structures and the aspects of the phagolysosomes. It was noteworthy the presence of giant phagolysosomes enclosing bacteria in active proliferation commonly observed in CHS neutrophils after 45 min of phagocytosis, wich corresponded with the impaired bactericidal activity of these leukocytes. The present results suggest that phagocytosis may also be defective in CHS, and point out to the sensitivity of the fluorochrome phagocytosis assay and its application in clinical laboratories