Aislamiento de Neisseria meningitidis en muestra anorectal en hombre que tiene sexo con hombre con uretritis y sifilis primaria / Isolation of Neisseria meningitidis in anorectal specimen in a men who have sex with men with uretritis and primary syphilis

Resumen Neisseria meningitidis es una bacteria gramnegativa asociada frecuentemente a enfermedades invasoras de elevada mortalidad. Si bien su reservorio natural es la nasofaringe humana, en los últimos años han aumentado los aislamientos de este agente en la mucosa anorectal, principalmente en hombres que tienen sexo con hombres (HSH). Presentamos el caso de un HSH con infección por VIH, que consultó por un cuadro de uretritis y sifilis primaria, en el cual se aisló N. meningitidis en una muestra anorectal. Fue tratado en forma empírica con ceftriaxona y azitromicina, realizándose un cultivo de control post-tratamiento que fue negativo. A pesar del aumento de las infecciones y colonizaciones anogenitales por N. meningitidis, se desconoce su rol como patógeno genital, en la transmisión de otras infecciones y la necesidad de esquemas terapéuticos específicos.

Abstract Neisseria meningitidis is a Gram-negative bacterium frequently associated with invasive diseases with high mortality. Although its natural reservoir is the human nasopharynx, in recent years there have been increasing reports of isolation of this agent in the anorectal mucosa, mainly in men who have sex with men (MSM). We present the case of an HIV-positive MSM who consulted for urethritis and primary syphilis, in which N. meningitidis was isolated in an anorectal specimen. He was treated empirically with ceftriaxone and azithromycin, and a post-treatment control culture was negative. Despite the increase in anogenital infections and colonization by N. meningitidis, its role is unknown as a genital pathogen and in the transmission of other infections and the need for specific therapeutic regimens.

Neisseria meningitidis aislada de muestras de hombres que tienen sexo con hombres / Neisseria meningitidis isolated from patients in men who have sex with men

Resumen En los períodos 2000-2004 y 2014-2015 se investigó la presencia de Neisseria meningitidis en 1.143 y 544 hombres que tienen sexo con hombres respectivamente, atendidos en el marco de un programa de enfermedades de transmisión sexual. Se determinó la prevalencia de este agente, su distribución en serogrupos y su sensibilidad a los antimicrobianos. Se obtuvieron hisopados faríngeos, rectales y uretrales, que se sembraron en medio selectivo Thayer Martin modificado. La identificación se realizó mediante pruebas bioquímicas convencionales y por espectrometría de masas (MALDI-TOF). En el segundo período estudiado, sobre 85 aislamientos procedentes de faringes se investigaron los serogrupos B, C, W e Y mediante PCR. Se determinó la CIM de penicilina, ceftriaxona, rifampicina, azitromicina y ciprofloxacina en 66 aislamientos obtenidos en el primer período y en 102 logrados en el segundo. La prevalencia de N. meningitidis fue del 17,8% en el primer período y del 28,1% en el segundo; este microorganismo se aisló más frecuentemente de fauces. Los serogrupos hallados fueron B (31,5%), Y (7,6%) y W (3,3%), con un 9,8% de aislamientos no capsulados; los restantes corresponderían a otros serogrupos. El 34,8% y el 63,7% de los aislados estudiados correspondientes al primer y segundo período, respectivamente, tuvieron sensibilidad intermedia a la penicilina, y un 11,8% de los evaluados en el segundo período fueron resistentes a dicho antibiótico. Todos los aislados estudiados fueron sensibles a ceftriaxona y a ciprofloxacina (excepto 3, con CIM entre 0,25 y 0,5(g/ml), el 3% fueron resistentes a rifampicina y el 2% fueron no sensibles a azitromicina. La portación de N. meningitidis en hombres que tienen sexo con hombres fue elevada y hubo un alto porcentaje de cepas no sensibles a penicilina. El serogrupo B fue prevalente.

Abstract During the periods 2000-2004 and 2014-2015, Neisseria meningitidis was investigated in men who have sex with men, 1143 and 544 respectively, who consulted in the sexually-transmitted disease program. Prevalence, serogroup distribution and susceptibility to antibiotics were determined. Pharyngeal, rectal and urethral swabs were cultivated on selective Thayer-Martin modified medium. The identification was performed by biochemical tests and mass spectrometry by MALDI-TOF. Serogroups B, C, W and Y were investigated by PCR in 85 isolates recovered from the pharynx belonging to the second period. MICs of penicillin, ceftriaxone, rifampicin, azithromycin and ciprofloxacin were determined for 66 and 102 isolates from periods 1 and 2 respectively, according to CLSI. The prevalence of N. meningitidis was 17.8% and 28.1%, in periods 1 and 2 respectively; the isolates were mainly recovered from the pharynx. The distribution of serogroups was B 31.5%; Y 7.6%; W 3.3% and 9.8% non-capsulated and the rest would belong to other serogroups. Isolates classified as intermediate to penicillin were 34.8% and 63.7% (first and second periods, respectively); moreover, 11.8% of the isolates from the second period were resistant. All isolates were susceptible to ceftriaxone, to ciprofloxacin (except 3 isolates with MIC values between 0.25 and 0.5(g/ml), 3% were resistant to rifampicin and 2% were not susceptible to azithromicin. The prevalence of N. meningitidis carriage in men who have sex with men was high with a high rate of penicillin non-susceptible isolates. B was the prevalent serogroup.

Genotipos de Neisseria meningitidis aislados de pacientes con enfermedad meningocócica en Paraguay, 1996-2015 / Genotypes of Neisseria meningitidis isolates in patients with meningococcal meningitis in Paraguay, 1996-2015 / Genótipos de Neisseria meningitidis isolados de pacientes com doença meningocócica no Paraguai, 1996-2015

RESUMEN Objetivo Describir las características fenotípicas y genotípicas de cepas de Neisseria meningitidis aisladas de enfermedad meningocócica en Paraguay entre 1996 y 2015. Métodos Se estudiaron por métodos microbiológicos convencionales y técnicas moleculares 114 aislamientos de N. meningitidis y 12 muestras clínicas sin aislamiento confirmadas por reacción en cadena de la polimerasa (PCR) que fueron remitidas por los diferentes centros centinelas y centros colaboradores de Paraguay. Resultados El grupo de edad más afectado fue el de menores de 1 año (19,0%), seguido por el de 1 a 5 años (17,5%). Un mayor porcentaje de las cepas se aisló de casos de meningitis (81,7%) y el serogrupo B se encontró en 60,3% de los casos. Los fenotipos más frecuentes fueron B:4:P1.14 (16,0%), B:15:P1.5, C:NT:NST y W:NT:P1.2 (10,5%), respectivamente. Los complejos clonales prevalentes fueron ST-11/ET37 complex 29,6% (8/27) con predominio del serogrupo W (6/8), ST-35 complex 18,5% (5/27) en el serogrupo B (4/4), y ST-32/ET5 complex 14,8% (4/16) en el serogrupo B (5/5). Conclusiones En Paraguay la enfermedad meningocócica es relativamente infrecuente. Los análisis de distribución de serogrupo muestran que el más frecuente es el B y en los últimos dos años aumentaron los casos de enfermedad meningocócica por C y W. Los complejos clonales encontrados se correlacionan con los hallados en la región del Cono Sur. Debido al alto nivel de virulencia de N. meningitidis, su vigilancia debe constituir una prioridad estratégica de los sistemas de salud pública nacionales y regionales para prevenir brotes epidémicos y apoyar la toma de decisiones en salud pública.

ABSTRACT Objective Describe the phenotypical and genotypical characteristics of Neisseria meningitidis isolates from cases of meningococcal disease in Paraguay between 1996 and 2015. Methods Conventional microbiological methods and molecular techniques were used to study 114 isolates of N. meningitidis and 12 clinical samples without isolation (confirmed by polymerase chain reaction), provided by various sentinel centers and collaborating centers in Paraguay. Results The most affected age group was children under 1 year (19.0%), followed by 1-5-year-olds (17.5%). The highest percentage of strains was isolated in meningitis cases (81.7%) and serogroup B was found in 60.3% of cases. The most frequent phenotypes were B:4:p1.14 (16.0%), B:15:p1.5, C:nt:nst, and W:nt:p1.2 (10.5%), respectively. The prevalent clonal complexes were: ST-11/ET37 complex, 29.6% (8/27), predominantly serogroup W (6/8); ST-35 complex, 18.5% (5/27), in serogroup B (4/4); and ST-32/ET5 complex, 14.8% (4/16), in serogroup B (5/5). Conclusions Meningococcal meningitis is relatively uncommon in Paraguay. Distribution analysis showed that serogroup B is the most common and that the number of cases of meningococcal disease caused by serogroups C and W increased in the last two years. The identified clonal complexes were correlated with those found in the Southern Cone region. Due to the high virulence of N. meningitidis, its surveillance should be a strategic priority of national and regional public health systems to prevent epidemic outbreaks and support public health decision-making.

RESUMO Objetivo Descrever as características fenotípicas e genotípicas de cepas de Neisseria meningitidis isoladas de casos de doença meningocócica no Paraguai entre 1996 e 2015. Métodos Foram estudados por métodos microbiológicos convencionais e técnicas moleculares 114 isolados de N. meningitidis e 12 amostras clínicas sem isolamento confirmadas por reação em cadeia da polimerase (PCR) enviados por diferentes centros-sentinela e centros colaboradores do Paraguai. Resultados A faixa etária mais afetada foi a de crianças menores de 1 ano (19,0%) e crianças de 1 a 5 anos (17,5%). Uma maior porcentagem de cepas foi isolada de casos de meningite (81,7%) e o sorogrupo B foi identificado em 60,3% dos casos. Os fenótipos mais comuns foram B:4:P1.14 (16,0%), B:15:P1.5, C:NT:NST e W:NT:P1.2 (10,5%), respectivamente. Os complexos clonais mais prevalentes foram o complexo ST-11/ET37 (29,6%, 8/27) com predomínio no sorogrupo W (6/8), complexo ST-35 (18,5%, 5/27) no sorogrupo B (4/4) e complexo ST-32/ET5 (14,8%, 4/16) no sorogrupo B (5/5). Conclusões A doença meningocócica é relativamente pouco comum no Paraguai. A análise da distribuição dos sorogrupos demonstrou que o sorogrupo B é o mais prevalente e, nos últimos dois anos, ouve um aumento nos casos de doença meningocócica pelos sorogrupos C e W. Os complexos clonais encontrados se correlacionam com os achados na região do Cone Sul. Devido à alta virulência da N. meningitidis, a vigilância deste agente deve ser uma prioridade estratégica dos sistemas de saúde pública nacionais e regionais para prevenir surtos epidêmicos e subsidiar a tomada de decisão em saúde pública.

Neumonía bacterémica por Neisseria meningitidis: primer caso reportado en Chile y revisión bibliográfica / Neisseria meningitidis pneumonia. a case report

Few cases of bacteremic pneumonia by Neisseria meningitidis (NM) have been described worldwide; mostly in elderly patients or those with comorbidities. They appear clinically indistinguishable from other acute infectious pneumoniae, that do not develope the syndrome of meningococcemia. We report a 17-years-old male, without prior medical history, consulting in the emergency department with a 7-day history of productive cough, right pleural pain, fever and dyspnea. He was admitted to the ICU due to septic shock and respiratory distress. He was managed with vasoactive drugs and prone positioning ventilation for 48 hours. Chest radiography showed a right superior lobe condensation. The electrocardiogram and echocardiogram suggested septic myocarditis. Blood cultures demonstrated the presence of serogroup W135-NM. A lumbar puncture ruled out meningitis, and a 10-day ceftriaxone therapy was completed favorably.

A cross-sectional study assessing the pharyngeal carriage of Neisseria meningitidis in subjects aged 1-24 years in the city of Embu das Artes, São Paulo, Brazil

ABSTRACT Meningococcal carriage is a prerequisite for invasive infection. This cross-sectional study assessed the pharyngeal carriage prevalence in healthy subjects aged 1-24 years in Embu das Artes city, São Paulo, Brazil. Pharyngeal swabs were examined for the presence of Neisseria meningitidis. The isolates were tested for different serogroups using agglutination and polymerase chain reaction. A logistic regression model assessed any independent association between Neisseria meningitidis carriage and various risk factors. A total of 87/967 subjects (9%, 95% Confidence Interval (CI): 7.3-11.0) tested positive for N. meningitidis: 6.2% (95% CI: 3.8-9.4) in 1-4 years, 8.5% (95% CI: 5.1-13.0) in 5-9 years, 12.5% (95% CI: 7.8-18.6) in 10-14 years, 12.6% (95% CI: 7.4-19.7) in 15-19 years and 9% (95% CI: 4.9-14.9) in 20-24 years age groups. Highest carriage prevalence was observed in adolescents 10-19 years old. Serogroup C was predominant (18.4%) followed by serogroup B (12.6%). The 15-19 years age group showed a significant association between number of household members and carriers of N. meningitidis. This cross-sectional study is the first in Brazil to evaluate meningococcal carriage prevalence and associated factors in a wide age range.

Caracterización clínica de la meningitis aguda de la comunidad causada por Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae y Listeria monocytogenes en adultos en el Hospital Carlos van Buren entre los años 2013 y 2016 / Clinical characteristics of community-acquired acute meningitis caused by Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae and Listeria monocytogenes in adults in Carlos van Buren Hospital between 2013-2016

INTRODUCCIÓN: la meningitis bacteriana es una enfermedad infecciosa aguda grave, que por su letalidad y costos en atención de salud genera un alto impacto en Salud Pública. Los agentes causales más frecuentes son Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae y Listeria monocytogenes, pero poco conocemos de nuestra realidad local. MATERIALES Y MÉTODOS: estudio descriptivo, con revisión de base de datos del laboratorio de microbiología del Hospital Carlos Van Buren, obteniendo datos de los cultivos de líquido céfalo raquídeo de pacientes mayores de 15 años entre marzo de 2013 y noviembre de 2016. RESULTADOS: 128 casos de meningitis bacteriana aguda, de los cuales 17 fueron por los microorganismos objetivos del estudio, siendo el más frecuente S. pneumoniae, clínicamente un 58% se presentó sin signos meníngeos. A 30 días del diagnóstico un 35% había fallecido, la mitad de ellos inició el tratamiento antibiótico pasadas las 24 horas desde su ingreso al hospital. En el 46% la tinción gram no evidenció bacterias. DISCUSIÓN: los microorganismos clásicamente descritos como agentes causales parecen no explicar la totalidad de los cuadros de meningitis bacteriana aguda en la población adulta estudiada, la ausencia de signos meníngeos no permite descartar la sospecha diagnóstica. La mitad de los pacientes fallecidos inició el tratamiento antibiótico pasadas las primeras 24 horas.

INTRODUCTION: bacterial meningitis is a serious acute infectious disease whose lethality and elevated health costs have a serious impact on public health. The most frequent causes are Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenza and Listeria monocytogenes, but we know little of the local situation. MATERIALS AND METHODS: A descriptive study reviewing Carlos van Buren Hospital´s microbiology laboratory data base, and obtaining the details of cerebrospinal fluid cultures of patients over the age of 15 between March 2013 and November 2016. RESULTS: 128 cases of acute bacterial meningitis of which 17% were caused by the micro-organisms of study, the most frequent being Streptococcus pneumoniae. 58% of patients had no meningeal signs. At 30 days from diagnosis 35% had died, half of these having started antibiotic treatment over 24 hours after admission. 46% of the Gram stains showed no bacteria. DISCUSSION: the classical infectious agents do not appear to account for the totality of acute bacterial meningitis in the population studied. The absence of meningeal signs should not rule out the diagnosis. Half of the patients who died started antibiotic treatment after the first 24 hours.

Comparison of culture and PCR methods in the diagnosis of bacterial meningitis

Abstract Our aim in this study is to compare the standard culture method with the multiplex PCR and the Speed-Oligo® Bacterial Meningitis Test (SO-BMT) – a hybridization-based molecular test method – during the CSF examination of the patients with the pre-diagnosis of acute bacterial meningitis. For the purposes of this study, patients with acute bacterial meningitis treated at the Dicle University Medical Faculty Hospital, Infectious Diseases and Clinical Microbiology Clinic between December 2009 and April 2012 were retrospectively evaluated. The diagnosis of bacterial meningitis was made based on the clinical findings, laboratory test anomalies, CSF analysis results, and the radiological images. Growth was observed in the CSF cultures of 10 out of the 57 patients included in the study (17.5%) and Streptococcus pneumoniae was isolated in all of them. The CSF samples of 34 patients (59.6%) were positive according to the SO-BMT and S. pneumoniae was detected in 33 of the samples (97.05%), while Neisseria meningitidis was found in 1 sample (2.95%). In a total of 10 patients, S. pneumoniae was both isolated in the CSF culture and detected in the SO-BMT. The culture and the SO-BMT were negative in 23 of the CSF samples. There was no sample in which the CSF culture was positive although the SO-BMT was negative. While SO-BMT seems to be a more efficient method than bacterial culturing to determine the pathogens that most commonly cause bacterial meningitis in adults, further studies conducted on larger populations are needed in order to assess its efficiency and uses.

Meningite aguda bacteriana na população infantojuvenil: principais agentes e métodos para diagnóstico - uma revisão da literatura / Acute bacterial meningitis in children and young population: main agents and methods for diagnosis - a literature review

A meningite bacteriana é uma afecção de grande significância devido a sua relação com alta mortalidade e morbidade na população neonatal a jovem. Devido a este fato é importante o conhecimento sobre esta doença e os seus principais agentes etiológicos. Com o objetivo de relatar os principais métodos de diagnóstico, assim como os principais agentes etiológicos envolvidos na fisiopatologia da meningite bacteriana em população jovem foi realizada uma busca por artigos publicados nos últimos 5 anos nas bases de dados Pubmed, Scielo, Bireme e Lilacs. A literatura atual aponta como microrganismos predominantes na incidência dessa doença a N. meningitidis S. pneumoniae, sendo as mais recorrentes na população de faixa etária entre 29 dias e 17 anos. Os fatores relacionados ao prognóstico estão intimamente relacionados com a distinção da classificação do agente etiológico em bacteriano ou viral, importante para a determinação da terapia adequada.

Bacterial meningitis is a highly significant disease due to its relationship with high mortality and morbidity in neonatal and young population. Due to this is important to know about this disease and its main etiological agents. In purpose to report the main diagnostic methods, as well as the main etiological agents involved in the pathophysiology of bacterial meningitis was conducted a search for articles published in the last five years in Pubmed, Scielo, Bireme and Lilacs. The current literature indicates N. meningitidis S. pneumoniae the most predominant microorganisms in the incidence of this disease between population aged between 29 days and 17 years. Factors related to prognosis are closely related to the classification distinction of the etiologic agent in bacterial or viral, important to determine the ap- propriate therapy.

Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples

Abstract Background Several in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans. Methods A total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae. Results All used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains. Conclusions The in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.

Detección de Neisseria meningitidis en portadores asintomáticos en un hospital universitario de Brasil / Detectio of Neisseria meningitidis in asymptomatic carriers in a university hospital from Brazil

Los portadores asintomáticos de meningococos en hospitales son un factor de riesgo (FR) para adquirir la enfermedad meningocócica. La frecuencia de portadores de meningococos fue determinada a través de colecta orofaríngea en personal de un hospital de Brasil (n = 200). La prevalencia de portadores fue del 9% (IC del 95%, 5-13%). Los FR asociados al estado de portador fueron los siguientes: edad promedio 26,5 años, sexo masculino, hábito de frecuentar bares y número de personas/casa. Entre las 18 cepas de meningococos aisladas, 14 eran no agrupables (NG), 3 correspondieron al serogrupo B y una al 29E. La frecuencia de los serotipos y serosubtipos fue heterogénea, con un ligero predominio de los serotipos 4 y 7 y de los serosubtipos P1.7 y P1.5. La mayoría de las cepas (n=13) fueron sensibles a los antimicrobianos estudiados. El gen ctrA fue identificado por PCR en 9 (64,3%) de las 14 cepas NG, lo que sugiere virulencia en la mayoría de las cepas NG aisladas. Por lo tanto, se requiere una vigilancia constante de estos portadores asintomáticos

Asymptomatic meningococcus carriers in hospitals is a risk factor for acquiring meningococcal disease. Meningococcal carrier (MC) frequency was investigated in oropharyngeal swab samples collected from 200 staff members at a teaching hospital from Brazil. MC prevalence was 9% (95% CI 5­13%). Risk factors associated with MC were: mean age of 26.5 years, male gender, bar attendance frequency and number of persons/house. Of 18 isolated meningococcal strains, 14 were non-groupable (NG), 3 corrresponded to serogroup B and 1 to serogroup 29E. The frequency of serotypes and serosubtypes was heterogenous, with a slight predominance of serotypes 4 and 7 and serosubtypes P1.7 and P1.5. Most strains (n=13) were susceptible to the antimicrobials tested. The ctrA gene (PCR) was identified in 9 (64.3%) of the 14 NG strains, suggesting virulence in most of the NG isolated strains. Therefore, a constant surveillance of these asymptomatic carriers is required

Etiologia e fatores prognósticos da sepse em crianças e adolescentes admitidos em terapia intensiva / Etiology and prognostic factors of sepsis among children and adolescents admitted to the intensive care unit

RESUMOObjetivo:Determinar a etiologia e as variáveis clínicas e evolutivas da sepse associadas ao prognóstico nos pacientes internados em unidade de terapia intensiva pediátrica.Métodos:Série de casos prospectiva e retrospectiva. Coleta de dados nos prontuários de pacientes com diagnóstico de sepse internados na unidade de terapia intensiva pediátrica de hospital geral, de janeiro de 2011 a dezembro de 2013. Foram identificadas bactérias em culturas de sangue e líquidos biológicos. As variáveis idade, sexo, esquema vacinal, comorbidades, uso prévio de antibióticos, dados clínicos à admissão e complicações na evolução foram comparadas nos grupos sobrevida e óbito (nível de significância de 5%).Resultados:Foram incluídos 115 pacientes, com média de idade de 30,5 meses. Etiologia bacteriana foi identificada em 40 pacientes. Perfusão periférica alterada à admissão e diagnóstico de sepse grave mostraram-se fatores associados às complicações. Houve maior número de complicações no grupo com idade maior de 36 meses (p = 0,003; odds ratio = 4,94). A presença de complicações durante a internação foi fator associado ao óbito (odds ratio = 27,7). As principais etiologias foram: bactérias Gram-negativas (15/40), Staphylococcus aureus (11/40) e Neisseria meningitidis (5/40).Conclusão:Bactérias Gram-negativas e Staphylococcus aureuspredominaram na etiologia da sepse em crianças e adolescentes admitidos em terapia intensiva. A gravidade da sepse e a perfusão periférica alterada à admissão estiveram associadas às complicações. A presença de complicações foi fator associado ao óbito.

ABSTRACTObjective:To determine the etiology and clinical disease progression variables of sepsis associated with the prognosis of patients admitted to a pediatric intensive care unit.Methods:Prospective and retrospective case series. Data were collected from the medical records of patients diagnosed with sepsis who were admitted to the pediatric intensive care unit of a general hospital from January 2011 to December 2013. Bacteria were identified in blood and fluid cultures. Age, sex, vaccination schedule, comorbidities, prior antibiotic use, clinical data on admission, and complications during disease progression were compared in the survival and death groups at a 5% significance level.Results:A total of 115 patients, with a mean age of 30.5 months, were included in the study. Bacterial etiology was identified in 40 patients. Altered peripheral perfusion on admission and diagnosis of severe sepsis were associated with complications. A greater number of complications occurred in the group of patients older than 36 months (p = 0.003; odds ratio = 4.94). The presence of complications during hospitalization was associated with death (odds ratio = 27.7). The main etiological agents were Gram-negative bacteria (15/40), Staphylococcus aureus (11/40) and Neisseria meningitidis (5/40).Conclusion:Gram-negative bacteria and Staphylococcus aureus predominated in the etiology of sepsis among children and adolescents admitted to an intensive care unit. The severity of sepsis and the presence of altered peripheral perfusion on admission were associated with complications. Moreover, the presence of complications was a factor associated with death.

nspA Gene as a specific genetic marker for detection of Neisseria meningitidis causing bacterial meningitis.

Bacterial meningitis caused by Neisseria meningitidis which causes human brain meninges damage, is generally diagnosed from patient cerebrospinal fluid through microscopy, immunological assays, biochemical test, PCR, microarray and biosensors. However, these methods are expensive, time-consuming or non-confirmatory due to certain limitations. A quick PCR based method was developed for detection of bacterial meningitis caused by N. meningitidis using specific primers based on amplification of virulence nspA (Neisseria surface protein A) gene partial sequence (202 bp). The nspA gene amplicon could be used as a genetic marker for minimum detection of 10 ng genomic DNA (G-DNA) of N. meningitidis with high sensitivity only in 80 min, which is least time reported for the confirmation of the disease. However, the lower detection limit was found as low as 1.0 ng G-DNA, but with less sensitivity. The cross-reactivity of the genetic marker was also studied with other possible pathogens. A comparison with the presently available detection methods and our method was also done using patient samples.

Epidemiological profile of meningococcal disease in the State of Minas Gerais and in the Central, North, and Triângulo Mineiro regions, Brazil, during 2000-2009 / Perfil epidemiológico da doença meningocócica, no Estado de Minas Gerais e nas regiões Centro, Norte e Triângulo Mineiro, Brasil, de 2000 a 2009

INTRODUCTION: Infection by Neisseria meningitidis, termed as meningococcal disease, can cause meningococcal meningitis and septicemia with or without meningitis. Meningococcal disease is endemic in Brazil and has a high potential to cause large-scale epidemics; therefore, it requires the immediate notification of cases to the Information System for Notifiable Diseases (SINAN) in Brazil. The aim of this study was to describe an epidemiological profile using data from notified and confirmed cases in the State of Minas Gerais, Brazil, from January 2000 to December 2009, obtained from the investigation records of individuals with meningitis registered with SINAN. METHODS: This was a retrospective, population-based study. Descriptive analysis of the data was made using the simple and relative frequencies of the categorical variables in the investigation records. RESULTS: There were 1,688 confirmed patients in Minas Gerais of which 45.5% lived in the Central, North, and Triângulo Mineiro regions. The highest frequencies of cases were in the 1-4-years age group (26.3%), males (54.7%), caucasian (36.4%), and lived in an urban area (80%). In the patients with specified education, 650 (60.9%) patients had secondary education. Serogrouping of meningococci had been performed in 500 (29.6%) patients by age and gender; 285 (57%) belonged to serogroup C, 67 (13.4%) were in the 1-to 4-years age group, and 168 (33.6%) were male. CONCLUSIONS: The epidemiological profiles of patients in the Central, North, and Triângulo Mineiro regions were not significantly different from the profile of patients in Minas Gerais.

INTRODUÇÃO: A infecção por Neisseria meningitidis, denominada doença meningocócica, pode causar meningite meningocócica e meningococcemia com ou sem meningite. A doença meningocócica é endêmica no Brasil; no entanto, o alto potencial de causar epidemias e letalidade elevada impõe a notificação imediata dos casos ao Sistema de Informação de Agravos de Notificação (SINAN) do Brasil. O objetivo deste estudo foi descrever o perfil epidemiológico, utilizando registros de casos notificados e confirmados no Estado de Minas Gerais, Brasil, de janeiro de 2000 a dezembro de 2009, obtidos a partir da ficha individual de investigação de meningites registrada no SINAN. M ÉTODOS: Trata-se de um estudo retrospectivo, de base populacional. Foram realizadas análises descritivas dos dados, a partir das frequências simples absolutas e relativas para as variáveis categóricas da ficha de investigação e teste qui-quadrado. RESULTADOS: Houve 1.688 casos confirmados em Minas Gerais, dos quais 45,5% residiam no centro, norte e Triângulo Mineiro. As maiores frequências foram nos pacientes do grupo etário de 1 a 4 anos (26,3%), sexo masculino (54,7%), brancos (36,4%) e residiam em zona urbana (80%). Dos casos com escolaridade especificada, 650 (60,9%) casos cursaram ensino médio completo. Foram sorogrupados e classificados segundo idade e sexo 500 (29,6%) casos, e desses 285 (57%) foram sorogrupo C, 67 (13,4%) da faixa etária de 1 a 4 anos e 168 (33,6%) do sexo masculino. CONCLUSÕES: O perfil epidemiológico dos casos nas regiões centro e norte e Triângulo Mineiro não apresentaram diferenças importantes comparado ao perfil dos casos de Minas Gerais.

Prevalence of Neisseria meningitidis carriers in children under five years of age and teenagers in certain populations of Mexico City / Prevalencia de portadores de Neisseria meningitidis en niños menores de cinco años de edad y adolescentes en determinadas poblaciones de la Ciudad de México

OBJECTIVE: To carry out a pilot study to discover the frequency of colonization in healthy children under five years old and teenagers, as well as the distribution of the different N. meningitidis serogroups isolated from nasopharyngeal samples collected from the population under study. MATERIAL AND METHODS: The population included youth between 15 and 19 years old living in social rehabilitation centers (SRC) and university teenagers (UT) as well as children under the age of five attending day care centers (DCC) in Mexico City. Nasopharyngeal exudates were processed using standard microbiological techniques in order to identify Nm, the serogroup, type and subtype of these isolates. RESULTS: A total of 2 310 samples (774 SRC, 800 UT and 736 DCC) were taken. Total prevalence of Nm was 1.6 per cent. In teenagers living in SRC the prevalence was 2.9 percent. The most frequent serogroups were Y (29.7 percent), C (24.3 percent) and B (10.8 percent). CONCLUSIONS: The prevalence of carriers in the study environment was significant (1.6 percent) and similar to countries in which the invasive illness is much more frequent.

OBJETIVO: Realizar un estudio piloto que permita conocer la frecuencia de colonización en niños sanos menores de cinco años y adolescentes, así como la distribución de serogrupos de los aislamientos de Neisseria meningitidis (Nm) obtenidos de la nasofaringe de la población estudiada. MATERIAL Y MÉTODOS: Se incluyeron, jóvenes entre los 15 y 19 años de edad, de centros de readaptación social (CRS) y adolescentes universitarios (AU), así como niños menores de cinco años que asisten a estancias infantiles (EI) en la Ciudad de México. Se tomaron exudados nasofaríngeos y fueron procesados, usando las técnicas microbiológicas clásicas, para identificar Nm, serogrupo, tipo y subtipo de estos aislamientos. RESULTADOS: Se incluyeron un total de 2310 muestras (774 CRS, 800 AU y 736 EI). La prevalencia total de Nm fue 1.6 por ciento y en los adolescentes de CRS fue 2.9 por ciento. Los serogrupos más frecuentes son; Y (29.7 por ciento), C (24.3 por ciento) y B (10.8 por ciento). CONCLUSIÓN: La prevalencia de portadores en nuestro medio fue significativa (1.6 por ciento) y similar a los países en los que la enfermedad invasiva es mucho más frecuente.

Caracterización de aislamientos invasivos de S. pneumoniae, H. influenzae y N. meningitidis en América Latina y el Caribe: SIREVA II, 2000-2005 / Characterization of invasive isolates of S. pneumoniae, H. influenzae, and N. meningitidis in Latin America and the Caribbean: SIREVA II, 2000-2005

OBJETIVOS: Analizar las características fenotípicas y la susceptibilidad a antibióticos de las cepas circulantes de Streptococcus pneumoniae, Haemophilus influenzae y Neisseria meningitidis en América Latina y el Caribe entre 2000 y 2005. Se evaluó la cobertura potencial de las vacunas conjugadas. MÉTODOS: Se estudió por métodos convencionales la distribución de los serotipos o serogrupos de 17 303 cepas de S. pneumoniae, 2 782 cepas de H. influenzae y 6 955 cepas de N. meningitidis aisladas de casos de neumonía, meningitis, sepsis, bacteriemias y otros procesos invasivos. Se evaluó la susceptibilidad a los antibióticos de las cepas estudiadas. Los aislamientos procedían de 453 centros centinelas de 19 países de América Latina y 4 del Caribe, como parte del proyecto SIREVA II. RESULTADOS: El serotipo 14 de S. pneumoniae fue el más frecuentemente aislado (21,1 por ciento), especialmente en niños menores de 6 años (29,1 por ciento). Las coberturas potenciales de las vacunas conjugadas antineumocócicas hepta, nona, deca y tridecavalentes fueron de 59,0 por ciento, 73,4 por ciento, 76,5 por ciento y 85,9 por ciento, respectivamente. De los aislamientos, 63,3 por ciento eran sensibles a la penicilina. El serotipo b de H. influenzae estuvo presente en 72,2 por ciento de los aislamientos en niños menores de 2 años, mientras 8,6 por ciento correspondieron a los serotipos a, c, d, e y f; 19,2 por ciento resultaron no serotipables. La proporción de cepas de H. influenzae productoras de betalactamasa en aislamientos en niños menores de 2 años fue de 16,3 por ciento. Los serogrupos de N. meningitidis más frecuentes fueron el B (69,0 por ciento) y el C (25,7 por ciento); 65,8 por ciento y 99,2 por ciento de las cepas fueron sensibles a la penicilina y a la rifampicina, respectivamente. CONCLUSIONES: Estos resultados resaltan la importancia de la vigilancia epidemiológica integral de S. pneumoniae, H. influenzae y N. meningitidis en América Latina...

OBJECTIVES: To analyze the phenotypical characteristics and the susceptibility to antibiotics of the circulating strains of Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis circulating in Latin America and the Caribbean from 2000-2005. Potential coverage by conjugate vaccines was evaluated. METHODS: Conventional methods were used to study the distribution of the serotypes or serogroups of 17 303 strains of S. pneumoniae, 2 782 strains of H. influenzae, and 6 955 strains of N. meningitidis isolated from cases of pneumonia, meningitis, sepsis, bacteriemias, and other invasive processes. The antimicrobial susceptibilities of the study strains were evaluated. The isolates came from 453 sentinel surveillance sites in 19 countries in Latin America and four in the Caribbean, as part of the SIREVA II (Network Surveillance System for the Bacterial Agents Responsible for Pneumonia and Meningitis) project. RESULTS: S. pneumoniae serotype 14 was the most frequently isolated (21.1 percent), especially in children under 6 years of age (29.1 percent). The potential coverages by hepta-, nona-, deca-, and trideca-valent antipneumonia conjugate vaccines were 59.0 percent, 73.4 percent, 76.5 percent, and 85.9 percent, respectively. Of the isolates, 63.3 percent were sensitive to penicillin. H. influenzae serotype b was present in 72.2 percent of the isolations from children under 2 years of age, whereas 8.6 percent produced serotypes a, c, d, e, and f, and 19.2 percent could not be serotyped. The rate of H. influenzae beta-lactamase-producing strains isolated from children under 2 years of age was 16.3 percent. The most frequent N. meningitidis serogroups were B (69.0 percent) and C (25.7 percent); 65.8 percent and 99.2 percent of the strains were susceptible to penicillin and rifampicin, respectively. CONCLUSIONS: These results highlight the importance of comprehensive epidemiological surveillance of S. pneumoniae, H. influenzae...

The utility of the polymerase chain reaction assay for aetiologic definition of unspecified bacterial meningitis cases

Most patients with acute suppurative meningitis are otherwise healthy individuals with regard to immune mechanisms against invasive bacterial disease. This medical emergency is among the most dramatic and potentially ravaging diseases that affect humans, particularly young children. The illness often strikes suddenly, and can either result in death or leave the survivors with significant neurological dysfunctions. The demonstration of a bacterial aetiology is necessary for decisions regarding treatment and prophylaxis. Conventional bacteriological methods frequently fail to identify an agent, as a result of administration of antibiotics or delayed lumbar punctures. We investigated the major aetiologic sources of unspecified bacterial meningitis cases (G00.9, ISCD-10) by polymerase chain reaction (PCR)-based identification of Neisseria meningitidis (crgA), Streptococcus pneumoniae (ply) and Haemophilus influenzae (bexA) in cerebrospinal fluid samples. The multiplex PCR detected N. meningitidis in 92 percent, S. pneumoniae in 4 percent and H. influenzae in 1 percent of the 192 clinical samples assayed; 3 percent were negative for all three DNA targets. Bacterial DNA detection was found to be a valuable adjunct to enhance bacterial meningitis surveillance when the yield of specimens by culture is reduced. The implementation of PCR assays as a diagnostic procedure in Public Health Laboratories is perceived to be a significant advance in the investigation of bacterial meningitis.

Diseño y evaluación de una reacción de polimerasa en cadena (RPC) múltiple, para la identificación de bacterias causantes de meningitis aguda en líquido cefalorraquídeo de niños chilenos / Multiplex PCR assay in spinal fluid to identify simultaneously bacterial pathogens associated to acute bacterial meningitis in Chilean children

Background: Acute bacterial meningitis (ABM) is a serious disease that needs rapid diagnosis for an accurate treatment. The most important etiological agents are: Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae type b. Overall pathogen detection rate in patients with ABM in Chile is 83 percent. Aim: To evaluate a Polymerase Chain Reaction (PCR) protocol for simultaneous detection of several pathogens in patients with ABM. Material and methods: We designed and evaluated a multiplex PCR protocol for simultaneous specific genes identifications of S pneumoniae (¡ytA and ply genes), N meningitidis (ctrA, crgA) and H influenzae (bexA) in cerebrospinal fluid (CSF) samples from pediatric patients with suspected diagnosis of ABM. Sensitivity, specificity and minimum detection levels of DNA were determined. Amplifications ofrDNA 16S gene was done to confirm extraction of bacterial DNA. Results: Ninety nine CSF samples were studied, 90 from children with fever and negative CSF culture, and 9 from ABM and positive culture patients. The PCR protocol had a sensitivity of 89 percent, specificity of 100 percent, positive predictive value 100 percent and negative predictive value 99 percent. Conclusions: We observed a high concordance (89 percent) between bacteriological cultures and the PCR protocol results. This diagnostic tool could increase identification of agents in specific settings such as patients previously treated with antibiotics.