RESUMO
Despite more than 50 years of vaccination, pertussis is still an endemic disease, with regular epidemic outbreaks. With the exception of Poland, European countries have replaced whole-cell vaccines (WCVs) by acellular vaccines (ACVs) in the 1990s. Worldwide, antigenic divergence in vaccine antigens has been found between vaccine strains and circulating strains. In this work, 466 Bordetella pertussis isolates collected in the period 1998-2012 from 13 European countries were characterised by multi-locus antigen sequence typing (MAST) of the pertussis toxin promoter (ptxP) and of the genes coding for proteins used in the ACVs: pertussis toxin (Ptx), pertactin (Prn), type 2 fimbriae (Fim2) and type 3 fimbriae (Fim3). Isolates were further characterised by fimbrial serotyping, multi-locus variable-number tandem repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE). The results showed a very similar B. pertussis population for 12 countries using ACVs, while Poland, which uses a WCV, was quite distinct, suggesting that ACVs and WCVs select for different B. pertussis populations. This study forms a baseline for future studies on the effect of vaccination programmes on B. pertussis populations.
Assuntos
Bordetella pertussis/classificação , Bordetella pertussis/isolamento & purificação , Variação Genética , Coqueluche/epidemiologia , Coqueluche/microbiologia , Antígenos de Bactérias/genética , Bordetella pertussis/genética , Eletroforese em Gel de Campo Pulsado , Europa (Continente)/epidemiologia , Humanos , Repetições Minissatélites , Epidemiologia Molecular , Tipagem de Sequências Multilocus , Toxina Pertussis/genética , Regiões Promotoras Genéticas , SorotipagemRESUMO
Pathogen adaptation has been proposed to contribute to the resurgence of pertussis. A striking recent example is the emergence of isolates deficient in the vaccine component pertactin (Prn). This study explores the emergence of such Prn-deficient isolates in six European countries. During 2007 to 2009, 0/83 isolates from the Netherlands, 0/18 from the United Kingdom, 0/17 Finland, 0/23 Denmark, 4/99 Sweden and 5/20 from Norway of the isolates collected were Prn-deficient. In the Netherlands and Sweden, respectively 4/146 and 1/8 were observed in a later period (201012). The Prn-deficient isolates were genetically diverse and different mutations were found to inactivate the prn gene. These are indications that Prn-deficiency is subject to positive selective pressure. We hypothesise that the switch from whole cell to acellular pertussis vaccines has affected the balance between 'costs and benefits' of Prn production by Bordetella pertussis to the extent that isolates that do not produce Prn are able to expand. The absence of Prn-deficient isolates in some countries may point to ways to prevent or delay the spread of Prn-deficient strains. In order to substantiate this hypothesis, trends in the European B. pertussis population should be monitored continuously.
Assuntos
Proteínas da Membrana Bacteriana Externa/análise , Proteínas da Membrana Bacteriana Externa/genética , Bordetella pertussis/isolamento & purificação , Fatores de Virulência de Bordetella/análise , Fatores de Virulência de Bordetella/genética , Coqueluche/prevenção & controle , Sequência de Aminoácidos , Sequência de Bases , Bordetella pertussis/genética , Criança , Pré-Escolar , Análise por Conglomerados , Doenças Transmissíveis Emergentes/genética , DNA Bacteriano/genética , Europa (Continente) , Feminino , Genótipo , Humanos , Lactente , Masculino , Tipagem Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA , Coqueluche/epidemiologia , Coqueluche/microbiologiaRESUMO
Despite extensive childhood immunisation, pertussis remains one of the world's leading causes of vaccine preventable deaths. The current methods used for laboratory diagnosis of pertussis include bacterial culture, polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) serology. We conducted a questionnaire survey to identify variations in the laboratory methods and protocols used among participating countries included in the European surveillance network for vaccine-preventable diseases(EUVAC.NET). In February 2010, we performed the survey using a web-based questionnaire and sent it to the country experts of 25 European Union countries,and two European Economic Area (EEA) countries,Norway and Iceland. The questionnaire consisted of 37 questions which covered both general information on surveillance methods and detailed laboratory methods used. A descriptive analysis was performed.Questionnaires were answered by all 27 contacted countries. Nineteen countries had pertussis reference laboratories at the national level; their functions varied from performing diagnosis to providing technical advice for routine microbiology laboratories. Culture,PCR and serology were used in 17, 18 and 20 countries,respectively. For PCR, nine laboratories used insertion sequence IS481 as the target gene, which is present in multiple copies in the Bordetella pertussis genome and thus has a greater sensitivity over single copy targets, but has been proved not to be specific for B.pertussis. Antibodies directed against pertussis toxin(PT) are specific for B. pertussis infections. For ELISA serology, only 13 countries' laboratories used purified PT as coating antigen and 10 included World Health Organization (WHO) or Food and Drug Administration (FDA) reference sera in their tests. This present survey shows that methods used for laboratory confirmation of pertussis differ widely among European countries and that there is a great heterogeneity of the reference laboratories and functions. To evaluate the effects of different pertussis immunisation programmes in Europe, standardisation and harmonisation of the laboratory methods are needed.
Assuntos
Bordetella pertussis/isolamento & purificação , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/normas , Métodos Epidemiológicos , Garantia da Qualidade dos Cuidados de Saúde/normas , Coqueluche/diagnóstico , Técnicas de Cultura , Ensaio de Imunoadsorção Enzimática , Europa (Continente) , Humanos , Internet , Reação em Cadeia da Polimerase , Vigilância de Evento Sentinela , Inquéritos e QuestionáriosRESUMO
Although previous studies using human cytokines in rabbits and rats have provided evidence of the participation of tumor necrosis factor alpha (TNF-alpha) and interleukin 1 beta (IL-1 beta) in the meningeal inflammatory cascade, the results obtained by several groups of investigators have been discordant or, at times, contradictory. In the present study, homologous cytokines were applied to the rabbit meningitis model. Intracisternal administration of 10(2)-10(5) IU of purified rabbit TNF-alpha (RaTNF-alpha) produced significant cerebrospinal fluid (CSF) inflammation. A similar response was observed after intracisternal inoculation of 5-200 ng of rabbit recombinant IL-1 beta (rrIL-1 beta). Preincubation of these two mediators with their specific antibodies resulted in an almost complete suppression of the CSF inflammatory response. In animals with Haemophilus influenzae type b lipooligosaccharide-induced meningitis, intracisternal administration of anti-rrIL-1 beta, anti-RaTNF-alpha, or both resulted in a significant modulation of meningeal inflammation. Simultaneous administration of 10(3) IU of RaTNF-alpha and 5 ng of rrIL-1 beta resulted in a synergistic inflammatory response manifested by a more rapid and significantly increased influx of white blood cells into the CSF compared with results after each cytokine given alone. These data provide evidence for a seminal role of TNF-alpha and IL-1 beta in the initial events of meningeal inflammation.
Assuntos
Infecções por Haemophilus/fisiopatologia , Interleucina-1/farmacologia , Meningite/fisiopatologia , Fator de Necrose Tumoral alfa/farmacologia , Animais , Anticorpos/administração & dosagem , Modelos Animais de Doenças , Haemophilus influenzae/patogenicidade , Imunização Passiva , Inflamação , Contagem de Leucócitos/efeitos dos fármacos , Lipopolissacarídeos/toxicidade , Coelhos , Proteínas Recombinantes/farmacologiaRESUMO
This study assessed the clinical value of routine follow-up chest radiographs in hospitalized children with community-acquired pneumonia. The study population consisted of 196 children hospitalized for community-acquired pneumonia diagnosed between 1993-1995. Seventeen infective agents (10 viruses and 7 bacteria) were sought. Chest radiographs were taken on admission and 3-7 weeks later. All children were treated with antibiotics. Data on the course of illness over the following 8-10 years were obtained from patient files and questionnaires sent to parents. A potential causative agent was found in 165 (84%) of 196 cases. On follow-up chest radiographs, residual or new changes were seen in 30% of cases. The residual changes tended to be more common after mixed viral-bacterial infection (43%) than after sole viral (25%) or sole bacterial (20%) infection. Interstitial infiltrates (66%), atelectasis (46%), and enlarged lymph nodes were the most common sequelae seen on follow-up. Residual findings on follow-up radiographs did not affect the treatment of the children. No further chest radiographs were taken. During the 8-10-year follow-up of 194 children, no illnesses appeared that were associated with previous pneumonia. Twenty-six children had a new episode of pneumonia, 7 of them had asthma, and 6 had different underlying illnesses. In conclusion, routine follow-up chest radiographs are not needed in childhood community-acquired pneumonia if the child has a clinically uneventful recovery.
Assuntos
Pneumonia/diagnóstico por imagem , Pneumonia/epidemiologia , Adolescente , Criança , Pré-Escolar , Infecções Comunitárias Adquiridas/diagnóstico por imagem , Infecções Comunitárias Adquiridas/epidemiologia , Infecções Comunitárias Adquiridas/terapia , Feminino , Finlândia/epidemiologia , Seguimentos , Humanos , Lactente , Recém-Nascido , Masculino , Avaliação de Processos e Resultados em Cuidados de Saúde , Pneumonia/terapia , Estudos Prospectivos , Radiografia Torácica/estatística & dados numéricos , RecidivaRESUMO
SETTING: Complications arising from bacille Calmette-Guérin (BCG) vaccination were recorded in a national register in Finland until 1988. In the period 1960-1988, 222 patients suffered from BCG osteitis. OBJECTIVE: To evaluate whether single nucleotide polymorphisms (SNPs) in the promoter region of the gene encoding interleukin 10 (IL-10) are associated with BCG osteitis after vaccination in neonates. DESIGN: Blood samples of 132 former BCG osteitis patients now aged 21-49 years were analysed in a controlled study for IL10 rs1800896 (-1082G/A), rs1800871 (-819C/T), rs1800872 (-592C/A) and rs1800890 (-3575T/A) polymorphisms. RESULTS: The frequencies of genotypes of IL10 rs1800896, rs1800871, rs1800872 and rs1800890, the frequencies of variant genotypes and the frequencies of major or minor alleles did not differ between patients and controls. Furthermore, the frequencies of the eight possible combinations of the three IL10 alleles located close to each other (IL10 rs1800896, IL10 rs1800871 and IL10 rs1800872) were surprisingly similar. CONCLUSION: Our results suggest that polymorphisms of the IL-10 encoding gene do not play a central role in the development of complications due to BCG vaccination, although the IL10 gene, especially IL10 rs1800896 (-1082G/A) polymorphism, is known to be associated with tuberculosis risk in Europeans and North Americans.
Assuntos
Vacina BCG/efeitos adversos , Interleucina-10/genética , Osteíte/induzido quimicamente , Adulto , Alelos , Vacina BCG/administração & dosagem , Estudos de Coortes , Feminino , Finlândia , Frequência do Gene , Genótipo , Humanos , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Osteíte/genética , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas , Estudos Prospectivos , Risco , Adulto JovemRESUMO
A monoclonal antibody (MAb)-based enzyme immunoassay was developed for detection of Haemophilus influenzae type b (Hib) lipooligosaccharides (LOS). The high affinity of polymyxin B for lipid A was used to bind the Hib LOS to microtiter wells. The immobilized LOS was detected with MAbs directed against the oligosaccharide component of Hib endotoxin. Hib LOS concentrations were measured in in vitro samples and in cerebrospinal fluid (CSF) sample obtained from rabbits with experimental Hib meningitis. The sensitivity of the assay was 1 ng LOS/ml sample and the results obtained with this assay correlated significantly with those obtained with the standard Limulus amebocyte lysate assay. This new assay provides a method for specific detection of Hib LOS in CSF samples and in aqueous laboratory fluids. This general methodology should also be useful for experimental research involving specific LPS/LOS molecules.
Assuntos
Anticorpos Monoclonais , Haemophilus influenzae/análise , Lipopolissacarídeos/análise , Polimixina B , Animais , Endotoxinas/análise , Lipopolissacarídeos/líquido cefalorraquidiano , Lipopolissacarídeos/imunologia , Polimixinas , CoelhosRESUMO
A sufficient sensitivity of PCR is a prerequisite for its use in the diagnosis of infectious diseases. We have used PCR for detecting gene elements of Borrelia burgdorferi, mycobacteria and Bordetella pertussis. With all these microbe groups, difficulties were encountered in achieving the demanded sensitivity with the primer pairs primarily selected. An extensive testing of various reaction parameters did not improve the sensitivity. Subsequently, we synthesized more primers derived from slightly different positions of the original target sequences. When the original and new primers were tested in possible combinations, some primer pairs reached 100-fold to 1000-fold higher sensitivity than the primary pairs. We conclude that in optimizing the sensitivity of PCR, more emphasis should be put on testing of several primer pairs than on the extensive screening of reaction parameters. Thus far, a trial-and-error approach has to be used, because there is no means to predict the sensitivity properties of a selected primer pair.
Assuntos
Primers do DNA/genética , Reação em Cadeia da Polimerase , Sequência de Bases , Bordetella pertussis/genética , Grupo Borrelia Burgdorferi/genética , Dados de Sequência Molecular , Mycobacterium tuberculosis/genética , Sensibilidade e EspecificidadeRESUMO
The clinical characteristics of acute otitis media in relation to coexisting respiratory virus infection were studied in a 1-year prospective study of 363 children with acute otitis media. Respiratory viruses were detected using virus isolation and virus antigen detection in nasopharyngeal specimens of 42% of the patients at the time of diagnosis. Rhinovirus (24%) and respiratory syncytial virus (13%) were the two most common viruses detected. Adenovirus, parainfluenza viruses, and coronavirus OC43 were found less frequently. The mean duration of preceding symptoms was 5.9 days before the diagnosis of acute otitis media. Ninety-four percent of the children had symptoms of upper respiratory tract infection. Fever was reported in 55% and earache in 47% of cases. Patients with respiratory syncytial virus infection had fever, cough, and vomiting significantly more often than patients with rhinovirus infection or virus-negative patients. No significant differences were found in the appearance of the tympanic membrane and outcome of illness between virus-negative and virus-positive patients with acute otitis. Most patients respond well to antimicrobial therapy despite the coexisting viral infection. If the symptoms of infection persist, they can be due to the underlying viral infection, and viral diagnostics preferably with rapid methods may be clinically useful in these patients.
Assuntos
Otite Média/etiologia , Infecções Respiratórias/complicações , Viroses/complicações , Doença Aguda , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Moraxella catarrhalis/isolamento & purificação , Otite Média/microbiologia , Estudos Prospectivos , Vírus Sinciciais Respiratórios/isolamento & purificação , Infecções Respiratórias/microbiologia , Rhinovirus/isolamento & purificaçãoRESUMO
OBJECTIVE: To assess the clinical characteristics of bacteremic urinary tract infection (UTI) in children. DESIGN: Clinical data of Finnish children with bacteremic UTI (n = 134) from 1985 to 1994 were analyzed. Their symptoms, laboratory and imaging findings were compared with those of age- and sex-matched patients hospitalized for blood culture negative UTI. RESULTS: Generally, no major differences were seen in clinical findings between bacteremic and nonbacteremic patients. Bacteremic children had more frequent feeding problems (P = 0.02), and children > or =12 months of age tended more often to have abdominal pain and vomiting than did nonbacteremic patients. Fever was the major initial symptom in both study groups, but no significant difference occurred in the mean highest temperature or in the mean of duration of fever before admission to the hospital. The mean concentration of serum C-reactive protein on admission was significantly higher in bacteremic patients (116 vs. 76 mg/l; P < 0.01). After onset of antimicrobial treatment fever lasted significantly longer in bacteremic patients than in control patients (means, 2.3 vs. 1.1 days; P < 0.01). Anatomic or functional abnormalities in the urinary tract were detected in 51% vs. 46%, respectively. Obstruction of the urinary tract (9% vs. 1%, P < 0.01) and Grade 3 to 5 vesicoureteral reflux (30% vs. 16%, P = 0.02) were significantly more frequent in bacteremic patients with UTI. Obstruction or vesicoureteral reflux was found in 46% of children with bacteremic UTI caused by Escherichia coli vs. 89% of children with non-E. coli infection (P < 0.01). CONCLUSIONS: Clinical symptoms do not significantly distinguish bacteremic from nonbacteremic children with UTI. Outcome of bacteremic UTI was comparable with that of nonbacteremic UTI. Bacteremic children, especially those with non-E. coli UTI, more often had anatomical or functional abnormalities in the urinary tract.
Assuntos
Infecções Bacterianas/patologia , Infecções Urinárias/patologia , Dor Abdominal/patologia , Infecções Bacterianas/epidemiologia , Infecções Bacterianas/microbiologia , Proteína C-Reativa/análise , Criança , Pré-Escolar , Escherichia coli/isolamento & purificação , Feminino , Febre/patologia , Finlândia/epidemiologia , Humanos , Lactente , Recém-Nascido , Masculino , Obstrução Ureteral/patologia , Infecções Urinárias/epidemiologia , Infecções Urinárias/microbiologia , Vômito/patologiaRESUMO
We studied the association of acute otitis media with different respiratory virus infections in a pediatric department on the basis of epidemics between 1980 and 1985. Altogether 4524 cases of acute otitis media were diagnosed. The diagnosis was confirmed by tympanocentesis in 3332 ears. Respiratory virus infection was diagnosed during the same period in 989 patients by detecting viral antigen in nasopharyngeal mucus. There was a significant correlation between acute otitis media and respiratory virus epidemics, especially respiratory syncytial virus epidemics. There was no significant correlation between outbreaks of other respiratory viruses and acute otitis media. Acute otitis media was diagnosed in 57% of respiratory syncytial virus, 35% of influenza A virus, 33% of parainfluenza type 3 virus, 30% of adenovirus, 28% of parainfluenza type 1 virus, 18% of influenza B virus and 10% of parainfluenza type 2 virus infections. These observations show a clear association of respiratory virus infections with acute otitis media. In this study on hospitalized children Haemophilus influenzae strains were the most common bacteriologic pathogens in middle ear fluid, occurring in 19% of cases. Streptococcus pneumoniae was present in 16% and Branhamella catarrhalis in 7% of cases. There was no association between specific viruses and bacteria observed in this study.
Assuntos
Otite Média/complicações , Infecções Respiratórias/complicações , Feminino , Finlândia , Haemophilus influenzae/isolamento & purificação , Humanos , Masculino , Moraxella catarrhalis/isolamento & purificação , Muco/microbiologia , Nasofaringe/microbiologia , Otite Média/epidemiologia , Otite Média/microbiologia , Vírus Sinciciais Respiratórios/isolamento & purificação , Infecções Respiratórias/epidemiologia , Infecções por Respirovirus/complicações , Streptococcus pneumoniae/isolamento & purificaçãoRESUMO
OBJECTIVE: To describe the characteristics of serologically diagnosed pneumococcal pneumonia and compare them with those of respiratory syncytial virus (RSV) pneumonia and bacteremic pneumococcal pneumonia. METHODS: IgG antibodies to pneumococcal pneumolysin and C-polysaccharide as well as immune complexes containing IgG antibodies to pneumolysin and C-polysaccharide were measured from acute and convalescent sera of 254 children with community-acquired pneumonia. Evidence of pneumococcal infection was found in 93 children. Clinical and laboratory data were retrospectively collected from the records of 38 children with sole (all tests for 16 other microbes negative) pneumococcal pneumonia and compared with 26 sole RSV-induced pneumonia from the present series and with the data of our 85 bacteremic pneumococcal pneumonia cases reported earlier. RESULTS: Serologically diagnosed sole pneumococcal pneumonia clinically overlapped with RSV pneumonia, but RSV pneumonia was more often associated with tachypnea (45% vs. 17%, P < 0.05) and low white blood cell counts (means, 12.0 x 109/l vs. 20.8 x 109/l; P < 0.001) as well as low serum C-reactive protein levels (means, 28 mg/l vs. 137 mg/l; P < 0.001). Alveolar infiltrates were found in 15% of chest radiographs of children with RSV pneumonia compared with 76% of those in children with sole pneumococcal pneumonia (P < 0.001). Patients with bacteremic pneumonia more often appeared ill (79% vs. 50%, P < 0.001) and more often had typical pneumococcal pneumonia with high fever, leukocytosis and lobar infiltrates in their chest radiographs (70% vs. 34%, P < 0.05) than those with serologically diagnosed pneumococcal pneumonia. CONCLUSIONS: Serologically detected pneumococcal pneumonia differs significantly from RSV pneumonia in laboratory and chest radiography findings, but the clinical signs and symptoms overlap considerably. Bacteremic pneumococcal pneumonia is a more severe illness than the serologically diagnosed one.
Assuntos
Bacteriemia/diagnóstico , Pneumonia Pneumocócica/sangue , Pneumonia Pneumocócica/diagnóstico , Infecções por Vírus Respiratório Sincicial/diagnóstico , Adolescente , Complexo Antígeno-Anticorpo/sangue , Complexo Antígeno-Anticorpo/imunologia , Bacteriemia/sangue , Bacteriemia/imunologia , Proteínas de Bactérias , Sedimentação Sanguínea , Proteína C-Reativa/análise , Criança , Pré-Escolar , Diagnóstico Diferencial , Feminino , Humanos , Imunoglobulina G/sangue , Lactente , Contagem de Leucócitos , Pulmão/diagnóstico por imagem , Pulmão/microbiologia , Pulmão/virologia , Masculino , Pneumonia Pneumocócica/imunologia , Polissacarídeos Bacterianos/imunologia , Radiografia , Infecções por Vírus Respiratório Sincicial/sangue , Infecções por Vírus Respiratório Sincicial/imunologia , Vírus Sinciciais Respiratórios/isolamento & purificação , Estudos Retrospectivos , Índice de Gravidade de Doença , Streptococcus pneumoniae/imunologia , Streptococcus pneumoniae/isolamento & purificação , Estreptolisinas/imunologiaRESUMO
OBJECTIVE: Serum procalcitonin (PCT), C-reactive protein (CRP) and interleukin-6 (IL-6) concentrations were measured in 126 children hospitalized for community-acquired, radiologically confirmed pneumonia to assess whether these host response values could be used to distinguish bacterial from viral pneumonia. METHODS: The samples for PCT, CRP and IL-6 measurements were obtained on admission or the first day of hospitalization. The etiology of pneumonia was studied with an extensive panel of methods that detected 6 bacteria and 11 viruses. RESULTS: In all, 54% had evidence of bacterial pneumonia, and 32% had evidence of sole viral pneumonia. In 14% of the cases the etiology could not be determined. Children with bacterial pneumonia had significantly higher PCT (median 2.09 ng/ml vs. 0.56 ng/ml, P = 0.019) and CRP concentrations (96 mg/l vs. 54 mg/l, P = 0.008) than those with sole viral etiology. However, the values markedly overlapped. No significant difference in IL-6 concentrations was seen between the two patient groups. Using PCT > or = 2.0 ng/ml, CRP > or = 150 mg/l or IL-6 > or = 40 pg/ml, the specificity was > or =80% for bacterial pneumonia. The sensitivities with these cutoff values were 50% for PCT, 31% for CRP and 34% for IL-6. CONCLUSIONS: The results indicate that the measurement of serum PCT, CRP and IL-6 has little value in the differentiation of bacterial and viral pneumonia in children. However, in some patients with very high serum PCT, CRP or IL-6 values, bacterial pneumonia is probable.
Assuntos
Proteína C-Reativa/análise , Calcitonina/sangue , Interleucina-6/sangue , Pneumonia Bacteriana/diagnóstico , Pneumonia Viral/diagnóstico , Precursores de Proteínas/sangue , Biomarcadores/sangue , Peptídeo Relacionado com Gene de Calcitonina , Criança , Pré-Escolar , Infecções Comunitárias Adquiridas/sangue , Infecções Comunitárias Adquiridas/diagnóstico , Diagnóstico Diferencial , Humanos , Pneumonia Bacteriana/sangue , Pneumonia Viral/sangue , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Pertussis is a highly contagious respiratory disease and the most serious effects occur in young infants. Recently it has been shown that rapid and highly specific PCR can be a useful diagnostic tool for detection of pertussis infection. To our knowledge there are no previous studies concerning the disappearance of Bordetella pertussis DNA from the nasopharynx during antimicrobial treatment. METHODS: We studied prospectively how rapidly live B. pertussis organisms and DNA of these bacteria disappear from the nasopharynx during erythromycin therapy in unvaccinated infants. Eighty-five nasopharyngeal swabs obtained from nine erythromycin-treated infants with pertussis on consecutive days during hospitalization were tested by PCR and culture. The PCR products were further analyzed by Southern hybridization. RESULTS: On the fourth day of treatment 56% of the samples were positive by culture and 89% by PCR, whereas after 7 days the rates were 0 and 56%, respectively. In seven of nine patients PCR remained positive for 1 to 7 days longer than culture. The follow-up study also showed the semiquantitative nature of the PCR assay. The intensity of the PCR products in agarose gel usually weakened with time during erythromycin therapy. CONCLUSIONS: The results of this study show that PCR assay can achieve the specific diagnosis of pertussis infection in a large proportion of infants even when antimicrobial treatment has killed the organisms and culture is no longer positive.
Assuntos
Técnicas Bacteriológicas , Bordetella pertussis/isolamento & purificação , Eritromicina/uso terapêutico , Nasofaringe/microbiologia , Reação em Cadeia da Polimerase , Coqueluche/diagnóstico , Sequência de Bases , Southern Blotting , Bordetella pertussis/efeitos dos fármacos , Bordetella pertussis/genética , DNA Bacteriano/análise , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Dados de Sequência Molecular , Estudos Prospectivos , Coqueluche/tratamento farmacológico , Coqueluche/microbiologiaRESUMO
BACKGROUND: Childhood community-acquired pneumonia is a common illness, but there have been relatively few comprehensive studies of the viral and bacterial etiology in developed countries. The aim of the present investigation was to determine the etiology of community-acquired pneumonia in hospitalized children by several laboratory methods. METHODS: In a 3-year prospective study a nasopharyngeal aspirate for viral studies and acute and convalescent serum samples for viral and bacterial serology were taken from 254 children with symptoms of acute infection and infiltrates compatible with pneumonia in the chest radiograph. The role of 17 microbes was investigated. RESULTS: A potential causative agent was detected in 215 (85%) of the 254 patients. Sixty-two percent of the patients had viral infection, 53% had bacterial infection and 30% had evidence of concomitant viral-bacterial infection. Streptococcus pneumoniae (37%), respiratory syncytial virus (29%) and rhinovirus (24%) were the most common agents associated with community-acquired pneumonia. Only one patient had a positive blood culture (S. pneumoniae) of 125 cultured. A dual viral infection was detected in 35 patients, and a dual bacterial infection was detected in 19 patients. CONCLUSIONS: The possible causative agent of childhood community-acquired pneumonia can be detected in most cases. Further studies are warranted to determine what etiologic investigations would aid in the management of pneumonia. With effective immunization for S. pneumoniae and respiratory syncytial virus infections, more than one-half of the pneumonia cases in this study could have been prevented.
Assuntos
Infecções Comunitárias Adquiridas/microbiologia , Pneumonia Bacteriana/microbiologia , Pneumonia Viral/microbiologia , Distribuição por Idade , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Infecções Comunitárias Adquiridas/diagnóstico , Infecções Comunitárias Adquiridas/epidemiologia , Feminino , Finlândia/epidemiologia , Hospitalização , Humanos , Incidência , Lactente , Masculino , Pneumonia Bacteriana/diagnóstico , Pneumonia Bacteriana/epidemiologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Estudos Prospectivos , Fatores de Risco , Estudos de Amostragem , Distribuição por SexoRESUMO
BACKGROUND: Increasing evidence suggests that cell-mediated immunity (CMI) is involved in immune response against Bordetella pertussis. However, there are practically no studies evaluating the significance of pertussis-specific CMI in relation to protection against clinical pertussis. METHODS: An outbreak of pertussis was studied prospectively in 13-year-old pupils in a rural school. B. pertussis infection was diagnosed by culture, microagglutination and enzyme immunoassay serology with the use of pertussis toxin, filamentous hemagglutinin and pertactin as antigens. Pertussis-specific CMI responses were assessed by in vitro proliferation assay of peripheral blood mononuclear cells. RESULTS: At the initial sampling 7 of 22 children had symptoms suggestive of pertussis and 15 were asymptomatic. Of the latter 3 remained healthy, 8 were later confirmed to have had asymptomatic infection, 3 developed laboratory-confirmed pertussis and 1 developed cough without laboratory evidence of pertussis. Initial in vitro proliferations of peripheral blood mononuclear cells induced by pertussis toxin, filamentous hemagglutinin and/or pertactin were positive in all 3 healthy children, in 6 of 8 children who had asymptomatic infection, but in none of the 3 children who later developed pertussis. Although some children who remained healthy had high values of antibodies, no clear association was found between initial serum antibody values and clinical outcome. CONCLUSIONS: These preliminary data suggest that CMI may have an important role in protection against clinical pertussis but do not exclude a role for antibodies. Furthermore the results stress a multifactorial nature of the immune protection against B. pertussis.
Assuntos
Anticorpos Antibacterianos/análise , Antígenos de Bactérias/análise , Bordetella pertussis/imunologia , Coqueluche/imunologia , Adolescente , Bordetella pertussis/isolamento & purificação , Surtos de Doenças , Feminino , Humanos , Imunidade Celular , Estudos Longitudinais , Masculino , Instituições Acadêmicas , Coqueluche/diagnóstico , Coqueluche/epidemiologia , Coqueluche/prevenção & controleRESUMO
Adjunctive dexamethasone therapy to antibiotic therapy for bacterial meningitis has been shown in several studies to be beneficial. The roles of interleukin 1 beta and tumour necrosis factor alpha (TNF-alpha) in bacterial meningitis and in an experimental rabbit model are reviewed; both cytokines play a pivotal role in induction of meningitis. Using dexamethasone with the initial dose of antibiotics caused a decrease in TNF-alpha and modulated the inflammatory response. Early use of adjunctive dexamethasone is important in therapy of bacterial meningitis.
Assuntos
Interleucina-1/fisiologia , Meningites Bacterianas/etiologia , Fator de Necrose Tumoral alfa/fisiologia , Animais , Criança , Dexametasona/uso terapêutico , Humanos , Meningites Bacterianas/tratamento farmacológico , Coelhos , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
During a local outbreak of pertussis mainly involving school-children and adults, 71 patients with suspected whooping cough and 25 of their household contacts were studied in the course of 1 day so as to compare enzyme-linked immunosorbent assay (ELISA) with bacterial isolation in the diagnosis of pertussis. Serum samples and two nasopharyngeal swabs were collected from all persons. Five different culture media were used. Twenty-eight cases were diagnosed within 2 days by measuring IgM- and IgA-class antibodies to Bordetella pertussis in the first serum samples of patients. Fifty cases were identified when paired serum samples were used. Cultures were positive in only nine patients. The results show that ELISA is a valuable aid in the diagnosis of pertussis in patients with negative cultures.
Assuntos
Coqueluche/diagnóstico , Adolescente , Anticorpos Antibacterianos/análise , Bordetella pertussis/isolamento & purificação , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina A/análise , Imunoglobulina M/análise , Lactente , Masculino , Nasofaringe/microbiologiaRESUMO
Major surgery, trauma, and infection induce a proinflammatory mediator response which, if excessive, may cause tissue injury. The response was measured during elective coronary bypass surgery when a centrifugal pump or a roller pump, differing in their basic working principles, was used for extracorporeal circulation (ECC). Eight patients were perfused with a centrifugal pump and eight patients with a roller pump during ECC. Plasma interleukin-1 beta (IL-1 beta), IL-2, IL-6, tumor necrosis factor alpha (TNF alpha), group II phospholipase A2, (PLA2), endotoxin, fibronectin and serum C-reactive protein (CRP) concentrations were measured. The operation increased plasma IL-6, group II PLA2, and serum CRP concentration and decreased plasma fibronectin concentrations. IL-1 beta and TNF alpha concentrations did not change. IL-2 occurred only occasionally, and endotoxin did not occur in any patient. No differences were seen between the group using a centrifugal pump and the group using the roller pump. Cardiac surgery with a perfusion time of less than two hours thus caused a proinflammatory mediator response which was similar whether a centrifugal or a roller pump was used for ECC.