Invasive non-type b haemophilus influenzae disease: report of eight cases / Doença invasiva causada por haemophilus influenzae não tipo b: relato de oito casos

ASTRACT Objective: To describe eight cases of invasive non-type b Haemophilus influenzae disease in children admitted to Hospital de Clínicas of Universidade Estadual de Campinas. Cases description: In 2015, there were eight cases of invasive non-type b H. influenzae disease. We tested the ampicillin sensitivity and beta-lactamase production of the strains identified and performed the genotyping. Molecular typing was determined by Pulsed-Field Gel Electrophoresis. Four patients were diagnosed with bacteremia; in two cases, H. influenzae was detected in the pleural fluid, and two patients had meningitis. Patients with comorbidities represented 37.5% of cases. Except for the strain of one patient - not sent to the reference laboratory -, all were ampicillin-sensitive and non-beta-lactamase-producing. Genotyping identified four non-capsular, one type c, and two type a strains. Molecular typing ruled out nosocomial transmission since all serotypes were distinct regarding genotype. Comments: The rise in cases of invasive non-type b H. influenzae infection was real. There was no nosocomial transmission, and we found no justification for the increase. These data indicate the need for surveillance to correctly diagnose, monitor, and understand the spectrum of non-type b H. influenzae disease.

ABSTRACT Objetivo: Descrever oito casos de doença invasiva por Haemophilus influenzae não tipo b em crianças internadas no Hospital de Clínicas da Universidade Estadual de Campinas. Descrição dos casos: Em 2015, ocorreram oito casos de doença invasiva por H. influenzae não tipo b. Nas cepas identificadas, testou-se a sensibilidade à ampicilina e a produção de betalactamase, e realizou-se a genotipagem. A tipagem molecular foi feita por Pulsed Field Gel Electrophoresis. Em quatro pacientes, o diagnóstico foi de bacteremia; em dois casos, H. influenzae foi identificado em líquido pleural, e dois pacientes tiveram meningite. Comorbidades foram encontradas em 37,5% dos pacientes. Com exceção da cepa de um dos pacientes (que não foi enviada ao laboratório de referência), todas eram sensíveis à ampicilina e não produtoras de betalactamase. A genotipagem identificou quatro cepas não capsulares, uma cepa tipo c e duas cepas tipo a. A tipagem molecular descartou a transmissão intra-hospitalar, já que todos os sorotipos eram distintos quanto ao genótipo. Comentários: O aumento dos casos de infecção invasiva por H. influenzae não tipo b foi real. Não houve transmissão intra-hospitalar e não foi encontrada justificativa para o aumento. Esses dados indicam a necessidade de vigilância para diagnosticar corretamente, monitorar e entender o espectro da doença causada por H. influenzae não tipo b.

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.

Meningite por Haemophilus influenzae tipo f / Meningitis due to Haemophilus influenzae type f

Com o declínio da taxa de infecções causadas pelo Haemophilus influenzae tipo b após a ampla introdução da vacina, sorotipos não-b devem ser considerados agentes patogênicos potenciais em crianças menores de 5 anos com doença invasiva. Relatamos um caso de meningite por Haemophilus influenzae tipo f em um lactente imunocompetente de 1 ano. O agente foi identificado em líquido cefalorraquidiano e hemocultura. A sorotipagem foi realizada por testes com soros policlonais e confirmada por reação em cadeia de polimerase. Todas as cepas de Haemophilus influenzae associadas à doença invasiva deveriam ser sorotipadas e notificadas, a fim de possibilitar uma análise adequada das mudanças e tendências na distribuição de sorotipos desta doença.

With the decline in the rate of infections caused by Haemophilus influenzae serotype b since the widespread vaccination, non-b serotypes should be considered as potential pathogenic agents in children with invasive disease younger than 5 years old. We report the case of an immunocompetent 1-year-old boy with Haemophilus influenzae type f meningitis. The agent was identified in cerebrospinal fluid and blood cultures. Serotyping was performed by tests using polyclonal sera and confirmed by polymerase chain reaction. All Haemophilus influenzae isolates associated with invasive disease should be serotyped and notified as a way to evaluate the changes and trends in serotype distribution of this disease.

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.

Simultaneous detection of Neisseria meningitidis, Haemophilus influenzae and Streptococcus sp. by polymerase chain reaction for the diagnosis of bacterial meningits

Avaliamos o desempenho da reação em cadeia da polimerase (PCR) para detecção simultânea da Neisseria meningitidis, Haemophilus influenzae e Streptococcus sp. no diagnóstico das meningites bacterianas e sua aplicabilidade na rotina diagnóstica. Foi realizado um estudo de coorte com 182 crianças apresentando suspeita de meningite bacteriana. Em 84, havia alterações clínicas e laboratoriais sugestivas de meningite bacteriana. Destas, 65 tiveram o agente etiológico identificado pelos métodos laboratoriais de rotina e 19 ficaram sem diagnóstico etiológico. Em 98 pacientes foi excluído o diagnóstico de meningite bacteriana. Analisando o desempenho da PCR encontramos sensibilidade de 88,1%, especificidade de 99,0% e valores preditivos positivo e negativo de 98,7% e 90,1% respectivamente. Nos 19 pacientes com meningite bacteriana mas sem diagnóstico etiológico a PCR detectou microrganismos em 14, sendo 12 N. meningitidis, um H. influenzae e um Streptococcus sp. A PCR possui o potencial de poder aumentar os índices de identificação das técnicas tradicionais, principalmente nas situações onde a microscopia direta, cultura ou identificação antigênica são negativos ou inconclusivos.

Haemophilus aphrophilus brain abscess in the first decade.

This report presents a case of brain abscess due to Haemophilus aphrophilus in a six-year old boy. He was admitted to our hospital suffering from left-sided weakness. The initial radiological diagnosis was an intracranial abscess. Purulent material was obtained by puncturing the subcortical lesion and the sample was cultured on conventional media. H. aphrophilus was isolated in pure culture, identified according to conventional methods and confirmed by Becton Dickinson Laboratory (BBL) crystal system. After surgical drainage and eight weeks of antibiotic therapy, the neurological findings improved. The presented case is an example of H. aphriphilus causing brain abscess in the first decade without cardiac predisposition and with good outcome.

Latex agglutination test in the diagnosis of pyogenic meningitis.

Samples of cerebrospinal fluid (n=204) from pediatric patients with clinically suspected pyogenic meningitis were examined by direct microscopy, bacterial culture and Latex Agglutination Test (LAT). Latex Agglutination Test was done for detection of antigen of Streptococcus pneumoniae and Haemophilus influenzae type b. Among 38 LAT positive cases, culture and/or gram stain was positive in only 20 cases and 18 cases were detected exclusively by LAT. Besides, LAT was useful in detecting the pre-treated cases as 11 out of 55 samples from pre-treated cases were positive by LAT in comparison to culture and/or Gram stain which detected only 4 of 55 cases. LAT is simple, rapid and more reliable test.

Disease burden and case management of bacterial meningitis among children under 5 years of age in Saudi Arabia

This study was designed to explore the case management and disease burden of bacterial meningitis among children below the age of 5 years in the Kingdom of Saudi Arabia. A prospective descriptive study was conducted during June 1999 through to May 2001 in 8 hospitals from 5 cities in different areas of the country. Those were, King Fahad Specialist Hospital and Maternity and Children Hospital, Buraidah, Belgorashi General Hospital and King Fahad Specialist Hospital, Al-Baha, Maternity and Children Hospital and Ohud Hospital, Al-Madina, Al-Qatif General Hospital in the Eastern Region and Prince Abdulrahman Bin Ahmed Alsudairy Central Hospital, Sakaka. The study population was 171,818 children under the age of 5 years. During the study period 208 cases of bacterial meningitis were identified, 141 [67.8%] with a definite causative organism: Hemophilus influenzae type b, Neisseria meningitidis, Streptococcus pneumoniae and other bacterial species. The remaining 67 cases [32%] were labeled as aseptic meningitis. A considerable proportion of cases was found to have received an antibiotic prior to presentation. While symptoms such as fever or poor feeding were common among cases, meningeal signs were less prominent. A lumbar puncture was carried out on all cases to reach diagnosis by gram stain, latex agglutination test and cerebrospinal fluid [CSF] and blood cultures following standard procedures. The immediate burden of meningitis cases was found to be the lengthy stay of patients in the hospital wards and intensive care units. Some of the main causative agents were resistant to the conventional antimicrobial therapy, but susceptible to newer antibiotics. The study being based on a population based surveillance gave a better overview on causative organisms of meningitis emphasizing that Gram stain, serology of CSF and culture [of CSF and blood] should be used. A high index of suspicion is needed to diagnose meningitis in children. Lumbar tap should be encouraged and supported in terms of training and more authorization to apply in diagnostic tests of such conditions. Audiometric measurement is a crucial need in the assessment of meningitis cases and in the process of their follow up. This type of service is clearly missing in our system. Influential steps are to be planned to avail this service

Meningitis and pneumonia in Guatemalan children: the importance of Haemophilus influenzae type b and Streptococcus pneumoniae

OBJECTIVE: To determine the epidemiology of Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae invasive infections in hospitalized Guatemalan children. This is an important issue since Hib vaccine has not been incorporated into the routine immunization program in Guatemala and information from hospital records in 1995 indicated a low incidence of Hib and S. pneumoniae as causes of meningitis and invasive infections. METHODS: Children who were hospitalized in Guatemala City with clinical signs compatible with bacterial infections were evaluated for evidence of Hib or S. pneumoniae infection. Normally sterile body fluids were cultured, and antigen detection was performed on cerebrospinal fluid (CSF) and pleural fluid. RESULTS: Of 1 203 children 1-59 months of age hospitalized over a 28-month period, 725 of them (60.3 percent) had a primary diagnosis of pneumonia, 357 (29.7 percent) of meningitis, 60 (5.0 percent) of cellulitis, and 61 (5.1 percent) of sepsis and other conditions. Hib was identified in 20.0 percent of children with meningitis and S. pneumoniae in 12.9 percent. The average annual incidence of Hib meningitis was 13.8 cases per 100 000 children under 5 years of age, and 32.4 percent of meningitides caused by Hib and 58.7 percent of S. pneumoniae meningitides occurred prior to 6 months of age. Case fatality rates were 14.1 percent, 37.0 percent, and 18.0 percent, respectively, for children with Hib, S. pneumoniae, and culture-negative and antigen-negative meningitis. Prior antibiotic therapy was common and was associated with significant reductions in CSF-culture-positive results for children with other evidence of Hib or S. pneumoniae meningitis. CONCLUSIONS: Improvements in case detection, culture methods, and latex agglutination for antigen detection in CSF resulted in identification of Hib and S. pneumoniae as important causes of severe disease in Guatemalan children. Using a cutoff of > 10 white blood cells per cubic millimeter in CSF would improve the sensitivity for detection of bacterial meningitis and help estimate the burden of bacterial meningitis in Guatemala and other developing countries.

Hemophilus influenzae serotype-b meningitis in an adult immunocompetent patient

We present a case of meningitis caused by Hemophilus influenzae type'b in an immunocompetent 41-year-old Saudi lady. The patient was successfully treated with Ceftriaxone for 10 days. A review of Hemophilus influenzae meningitis in adults and the impact of conjugated vaccine on the epidemiology of the disease are given

Rapid detection of Haemophilus influenzae type b in Bangladeshi children with pneumonia and meningitis by PCR and analysis of antimicrobial resistance.

A polymerase chain reaction (PCR) assay with primers from 'bexA' gene was compared with culture for the detection of Haemophilus influenzae type b (Hib) in clinical samples from children with pneumonia and meningitis. Of 200 sera (180 from pneumonia, 20 from non-pneumonia patients) tested by PCR (serum-PCR), Hib was detected in 15 of 16 blood culture-positive and in 6 blood culture-negative pneumonia cases. When compared with the results of blood culture, serum-PCR had sensitivity, specificity, and accuracy index of 93.7%, 96.7%, and 96.5% respectively. Of 120 cerebrospinal fluid (CSF) samples from meningitis patients tested by culture and PCR (CSF-PCR), the latter method could detect Hib in all 15 culture-positive and in 8 of 105 culture-negative cases, showing sensitivity, specificity, and accuracy index of 100%, 92.4%, and 94.4% respectively. The PCR result was available within a day. Antimicrobial susceptibility of Hib was determined by the disc-diffusion method. High rate of resistance to ampicillin (54.8%), chloramphenicol (48.4%), and co-trimoxazole (80.6%) was observed among 31 invasive Hib isolates with resistance to all 3 drugs (multiresistance) in 48.4% of the isolates. All the Hib isolates were susceptible to ceftriaxone. The study has shown that PCR is a rapid, sensitive and specific diagnostic test for Hib from clinical samples, and a combination of culture and PCR is necessary for the detection of Hib infections to the maximum extent for case management to reduce morbidity, mortality, and complications of the invasive Hib infections. A high prevalence of multiresistant Hib strains is a matter of concern.

Sindrome hemofagocitica secundaria / Secondary hemophagocytic syndrome

Embora consideradas doencas raras, as sindromes hemofagociticas representam verdadeiros desafios para o diagnostico, bem como para o tratamento. Os autores relatam o caso de uma crianca de 7 anos de idade, que desenvolveu a doenca apos uma meningite por Haemophilus influenzae e discutem os aspectos fisiopatologicos, clinicos e terapeuticos das sindromes hemofagociticas

Systemic Haemophilus influenzae disease in Thai children.

Fifty patients with systemic Haemophilus influenzae disease were indentified by hospital chart review between 1980-1992. The age distribution varied from 8 days to 14 years; the mean age of the patients was 12.7 months. The peak incidence was between 4 and 6 months of age. There were 27 male patients and 23 female patients for a male:female ratio of 1.17:1. The relative frequencies of 79 clinical entities encountered in 50 patients are as follows: meningitis 55.7%, bacteremia 13.9%, pneumonia 25.3%, cellulitis 2.5% arthritis 1.3%, septic shock 1.3%. There were 23 patients (46%) who had more than one disease entity. Most of the patients were anemic (Hb < 10 gm%) when hospitalization. Sixty-four percent of the patients had early complications. The mortality rate was 8%. Although serotyping was not done from the isolates, at least 33 cerebrospinal fluid samples were positive for H. influenzae type b capsular antigen by counterimmunoelectrophoresis. The percentage of susceptible H. influenzae to penicillin, ampicillin, chloramphenicol, co-trimoxazole were 57.1%, 76.4%, 87.5% and 54.2%, respectively. There was no strain resistant to third generation cephalosporin. Our data indicate that H. influenzae is a serious and life threatening infection. Early diagnosis and proper treatment will reduce the morbidity and mortality rates. For prevention of infection, an appropriate strategy for vaccination is required.

Meningitis neonatal por haemophylus / Neonatal meningitis by haemophylus

Se presenta el caso de un recién nacido con una meningitis connatal por haemophylus influenzae serotipo b, de transmisión vertical. Se destaca el hecho que este agente se aisla raramente en las infecciones neonatales, siendo ésta la primera comunicación en la literatura nacional. Se revisan las principales deiferencias que existen con los casos publicados en el extranjero

Contraimunoeletroforese no diagnóstico etiológico das meningites bacterianas / Counterimmunoelectrophoresis on etiological diagnosis of bacterial meningitis

Contraimunoeletroforese (CIE) tem sido utilizada para o diagnóstico etiológico de meningites bacterianas, através da detecçäo de antígenos polisacarídicos capsulares no líquido cefalorraquiano. O material é composto por 340 exames de amostras de LCR de 292 pacientes, 171 do sexo masculino e 121 do sexo feminino com idade entre 4 dias e 74 anos. Sendo 91 pacientes com perturbaçöes neurológicas diversas e 146 pacientes com diversas formas de meningites sépticas nas quais o agente etiológico foi identificado pelo exame bacterioscópico direto e provas culturais e 55 casos de meningites sépticas em que o agente etiológico näo foi identificado; em 25 pacientes foram feitos exames em 2 amostras; em 10, em 3 amostras e em 1, em 4 amostras. A primeira amostra foi sempre colhida antes da antibioticoterapia e as seguintes após o início da terapêutica. Este material foi dividido em grupos, para se verificar a especificidade e a sensibilidade da contraimunoeletroforese para Haemophilus influenzae tipo B, Neisseria meningitidis A, B e C e Streptococcus pneumoniae (83 sorotipos). A especificidade da contraimunoeletroforese para o Haemophilus influenzae é de 98, 49 e a sensibilidade é de 94,8%. Para o Neisseria meningitidis a especificidade éde 98,58% e a sensibilidade é de 44,4% e, para o Streptococcus pneumoniae a especificidade éde 93,55% e sensibilidade é de 41,1%.

Doença grave, diagnóstico precoce: meningite purulenta e teste de aglutinaçäo de partículas de látex / Grave disease, early diagnosis: bacterial meningitis and latex fixation tests

Desenvolveu-se um estudo transversal controlado prospectivo com o objetivo de avaliar, de forma independente e "cega" e em três etapas distintas e complementares, o desempenho do teste de aglutinaçäo de partículas de látex no diagnóstico etiológico das meningites determinadas pelo H. influenzae b, S. pneumoniae e N. meningitidis A e C em 284 crianças (casos e controles) com idade compreendida entre 3 meses e 14 anos. Na primeira, o teste em estudo foi comparado com um padräo ouro e revelou sensibilidade, especificidade, valor preditivo positivo, valor preditivo negativo e acurácia equivalentes a 92,7 por cento, 100,0 por cento, 95,0 por cento e 96,9 por cento, respectivamente. Na segunda estabeleceu-de uma comparaçäo com o exame quimiocitológico liquórico, quando entäo a sensibilidade, o valor preditivo negativo e a acurácia reduziram para 63,3 por cento, 70,8 por cento e 80,6 por cento, mantendo-se inalteradas as proporçöes da especificidade do valor preditivo positivo. Na última etapa, associou-se ao teste em estudo à bacterioscopia direta pelo Gram, confrontando-se ambos os métodos, com a cultura e contraimunoeletroforese, visando verificar o comportamento de cada dupla de exames no diagnóstico etiológico, independente do microorganismo causal. Näo ocorreu diferença estatisticamente significante na performance de cada uma dessas duplas de métodos laboratoriais (p=.79). Considerando que além do desempenho observado nesta investigaçäo, o teste em estudo reune outras vantagens como rapidez no fornecimento dos resultados e simplicidade de execuçäo e interpretaçäo, conclui-se que o mesmo representa um instrumento diagnóstico valioso tanto do ponto de vista clínico-individual como em programas de saúde coletiva.

Difusión de H. influenzae tipo B (HI B) en contactos intrafamiliares de niños con infecciones sistémicas por este agente / Spread of haemophilus type B in household contacts of patients with invasive infections

Actualmente está bien documentada la difusibilidad de Hi b especialmente en niños y se ha planteado quimioprofilaxis a los menores de 4 años que son contactos de infecciones sistémicas. Para conocer nuestra situación epidemiológica, se estudiaron 31 infecciones sistémicas por Hi b en pacientes ingresados a los hospitales L. Calvo Mackenna y Exequiel González Cortés (24 meningitis, 3 celulitis faciales, 1 neumonía abscesada, 3 pleuroneumonías, una de las cuales presentó además pericarditis) y se investigó la presencia de portadores entre los contactos intrafamiliares. Se estudiaron 191 contactos (X 6,4 por cada caso índice), encontrándose 18% de portadores entre los niños y 5,9% entre los adultos. Se detectó un caso secundario en un contacto de 7 meses de edad. Se logró la erradicación de todos los portadores mediante tratamiento con rifampicina durante 4 días. Se determinó el perfil de sensibilidad a los antimicrobianos de las cepas de Hi aisladas de pacientes y portadores, observándose 12% de cepas resistentes a ampicilina y cloramfenicol entre las cepas de los casos índices. No se observó resistencia en Hi b aislados de portadores