Antimicrobial resistance among invasive Haemophilus influenzae strains: results of a Brazilian study carried out from 1996 through 2000.

A total of 1712 strains of Haemophilus influenzae isolated from patients with invasive diseases were obtained from ten Brazilian states from 1996 to 2000. beta-Lactamase production was assessed and the minimum inhibitory concentrations (MIC) of ampicillin, chloramphenicol, ceftriaxone and rifampin were determined using a method for broth microdilution of Haemophilus test medium. The prevalence of strains producing beta-lactamase ranged from 6.6 to 57.7%, with an overall prevalence of 18.4%. High frequency of beta-lactamase-mediated ampicillin resistance was observed in Distrito Federal (25%), São Paulo (21.7%) and Paraná (18.5%). Of the 1712 strains analyzed, none was beta-lactamase negative, ampicillin resistant. A total of 16.8% of the strains were resistant to chloramphenicol, and 13.8% of these also presented resistance to ampicillin, and only 3.0% were resistant to chloramphenicol alone. All strains were susceptible to ceftriaxone and rifampin and the MIC90 were 0.015 micro g/ml and 0.25 micro g/ml, respectively. Ceftriaxone is the drug of choice for empirical treatment of bacterial meningitis in pediatric patients who have not been screened for drug susceptibility. The emergence of drug resistance is a serious challenge for the management of invasive H. influenzae disease, which emphasizes the fundamental role of laboratory-based surveillance for antimicrobial resistance.

Antimicrobial resistance among invasive Haemophilus influenzae strains: results of a Brazilian study carried out from 1996 through 2000

A total of 1712 strains of Haemophilus influenzae isolated from patients with invasive diseases were obtained from ten Brazilian states from 1996 to 2000. ß-Lactamase production was assessed and the minimum inhibitory concentrations (MIC) of ampicillin, chloramphenicol, ceftriaxone and rifampin were determined using a method for broth microdilution of Haemophilus test medium. The prevalence of strains producing ß-lactamase ranged from 6.6 to 57.7 percent, with an overall prevalence of 18.4 percent. High frequency of ß-lactamase-mediated ampicillin resistance was observed in Distrito Federal (25 percent), Säo Paulo (21.7 percent) and Paraná (18.5 percent). Of the 1712 strains analyzed, none was ß-lactamase negative, ampicillin resistant. A total of 16.8 percent of the strains were resistant to chloramphenicol, and 13.8 percent of these also presented resistance to ampicillin, and only 3.0 percent were resistant to chloramphenicol alone. All strains were susceptible to ceftriaxone and rifampin and the MIC90 were 0.015 æg/ml and 0.25 æg/ml, respectively. Ceftriaxone is the drug of choice for empirical treatment of bacterial meningitis in pediatric patients who have not been screened for drug susceptibility. The emergence of drug resistance is a serious challenge for the management of invasive H. influenzae disease, which emphasizes the fundamental role of laboratory-based surveillance for antimicrobial resistance

Immunodiagnosis of pneumococcal meningitis using Dot-enzyme-linked immunosorbent assay.

A diagnosis of bacterial meningitis requires isolation of the pathogen from cerebrospinal fluid (CSF). However, cultures of CSF are usually insensitive, thus, in the majority of patients, the etiology is rarely determined. A total of 90 CSF samples from pediatric patients with clinical diagnosis of bacterial meningitis were evaluated by Dot-ELISA. This method was standardized in order to detect pneumococcal polysaccharide antigen in CSF samples previously treated with 0.1 M EDTA and dotted on nitrocellulose membrane strips. Pneumococcal omniserum diluted 1:200 was employed for pneumococcal antigen detection. Dot-ELISA showed relative indices of 100 and 90 per cent for sensitivity and specificity, respectively. This method is cheaper than counter immunoelectrophoresis for pneumococcal antigen detection.

Detecting polysaccharide antigen of Neisseria meningitidis group C in cerebrospinal fluid by dot-ELISA assay.

Cerebrospinal fluid (CSF) samples from 210 patients (200 with clinical evidence of bacterial meningitis, 10 with other clinical neurologic disease) were tested by a Dot-ELISA assay for detection of polysaccharide antigen of N. meningitidis group C. CSF samples were treated with EDTA 0.1 M, at pH 7.5 and heated to 90>C for 10 min. Polyclonal antiserum was purified by use of ethanol fractionation. The results were compared to those using bacterial culture (BC), latex agglutination (LA), counterimmunoelectrophoresis (CIE), and direct microscopy (DM) methods. Test results showed a correlation of 93.3%, 94.3%, 91.0% and 69.5% respectively, and sensitivity of 0.947 and specificity of 0.930. This study suggests that the dot-ELISA assay of CSF is a useful alternative technique for the diagnosis of group C meningitis.

Antimicrobial resistance patterns of Haemophilus influenzae isolated from patients with meningitis in São Paulo, Brazil.

From 1989 to 1995, a total of 391 Haemophilus influenzae isolates were recovered from the cerebrospinal fluid (CSF) of hospitalized patients in São Paulo, Brazil. The majority of strains were isolated from infants aged less than 5 years. Strains belonging to biotype I (64.7%), biotype II (34.5%) and biotype IV (0.76%) were detected. Ninety-nine percent of these strains were serotype b. Minimal inhibitory concentration (MIC) was determined for ampicillin, chloramphenicol and ceftriaxone. The ss-lactamase assay was performed for all strains. The rate of ss-lactamase producer strains ranged from 10 to 21.4% during a period of 7 years, with an overall rate of 13.8%. Of the 391 strains analyzed, none was ss-lactamase negative ampicillin resistant (BLNAR). A total of 9.7% of strains showed resistance to both ampicillin and chloramphenicol; however, 4% of them were resistant to ampicillin only and 2% to chloramphenicol. All strains were susceptible to ceftriaxone and the MIC90 was 0.007 microg/ml, suggesting that ceftriaxone could be an option for the treatment of bacterial meningitis in pediatric patients who have not been screened for drug sensitivity.

Antimicrobial resistance patterns of Haemophilus influenzae isolated from patients with meningitis in São Paulo, Brazil

From 1989 to 1995, a total of 391 Haemophilus influenzae isolates were recovered from the cerebrospinal fluid (CSF) of hospitalized patients in São Paulo, Brazil. The majority of strains were isolated from infants aged less than 5 years. Strains belonging to biotype I (64.7 per cent), biotype II (34.5 per cent) and biotype IV (0.76 per cent) were detected. Ninety-nine percent of these strains were serotype b. Minimal inhibitory concentration (MIC) was determined for ampicillin, chloramphenicol and ceftriaxone. The ß-lactamase assay was performed for all strains. The rate of ß-lactamase producer strains ranged from 10 to 21.4 per cent during a period of 7 years, with an overall rate of 13.8 per cent. Of the 391 strains analyzed, none was ß-lactamase negative ampicillin resistant (BLNAR). A total of 9.7 per cent of strains showed resistance to both ampicillin and chloramphenicol; however, 4 per cent of them were resistant to ampicillin only and 2 per cent to chloramphenicol. All strains were susceptible to ceftriaxone and the MIC90 was 0.007 µg/ml, suggesting that ceftriaxone could be an option for the treatment of bacterial meningitis in pediatric patients who have not been screened for drug sensitivity.

[Detection of Neisseria meningitidis group B antigens by MB-Dot-Elisa test in patients with meningitis]. / Detección de antígenos de Neisseria meningitidis del grupo B con la prueba MB-Dot-ELISA en pacientes con meningitis.

Infection with Neisseria meningitidis group B has been difficult to detect, partly because this bacterial group's polysaccharide is a weak immunogen. This article describes work carried out to test a new procedure (MB-Dot-ELISA) employing a high-titered horse antiserum for detection of N. meningitidis group B antigens. The study assayed cerebrospinal fluid samples from 585 subjects, 574 with suspected meningitis cases and 11 with neurologic disorders. The results of the assay indicated a sensitivity of 0.991 and a specificity of 0.826. These results were superior to those obtained with latex agglutination and in substantial agreement with the results of counterimmunoelectrophoresis and bacteriologic methods. Overall, the MB-Dot-ELISA was found to be sensitive, inexpensive, and suitable for public health laboratory investigations.

Detection of N. meningitidis group B antigens by MB-Dot-ELISA in patients with meningitis.

Infection with Neisseria meningitidis group B has been difficult to detect, partly because this bacterial group's polysaccharide is a weak immunogen. This article describes work carried out to test a new procedure (MB-Dot-ELISA) employing a high-titered horse antiserum for detection of N. meningitidis group B antigens. The study assayed cerebrospinal fluid samples from 585 subjects, 574 with suspected meningitis cases and 11 with neurologic disorders. The results of the assay indicated a sensitivity of 0.991 and a specificity of 0.826. These results were superior to those obtained with latex agglutination and in substantial agreement with the results of counterimmunoelectrophoresis and bacteriologic methods. Overall, the MB-Dot-ELISA was found to be sensitive, inexpensive, and suitable for public health laboratory investigations.

Diagnosis of bacterial meningitis and septicemia by serum counterimmunoelectrophoresis.

We compare the results obtained by counterimmunoelectrophoresis in samples of serum and cerebrospinal fluid with microbiologic methods for 3,298 patients suspected of bacterial meningitis and/or septicemia at Instituto Adolfo Lutz, São Paulo, in a retrospective study of the period from July 1998 to July 1994. Of the 415 patients (12.6% of the total cases studied) who were positive by the serum test, only 249 (7.6% of the total cases studied) were also positive when cerebrospinal fluid was assayed. Thus, 40% of the positives (5.6% of the total) were identifiable by analysis of serum but not of cerebrospinal fluid. Neisseria meningitidis accounted for 77.7% (129) and Haemophilus influenzae for 22.3% (37) of the positive results obtained only when serum was examined. These data show that although sensitivity and specificity of serum counterimmunoelectrophoresis are relatively low compared to cerebrospinal fluid counterimmunoelectrophoresis, the serum test is necessary to complement cerebrospinal fluid counterimmunoelectrophoresis data.

Distribution of serotypes of Streptococcus pneumoniae isolated from invasive infections over a 16-year period in the greater São Paulo area, Brazil.

Capsular types of pneumococci from normally sterile body sites of 1,622 patients in Brazil were analyzed. Of 1,477 isolates from cerebrospinal fluid, 76.1% were of types represented in the currently available pneumococcal vaccine. The importance of age, time, and place in determining the optimal formulation of pneumococcal vaccine is considered.

Diagnosis of bacterial meningitis and septicemia by serum counterimmunoelectrophoresis

We compare the results obtained by counterimmunoelectrophoresis in samples of serum and cerebrospinal fluid with microbiologic methods for 3,298 patients suspected of bacterial meningitis and/or septicemia at Instituto Adolfo Lutz, Säo Paulo, in a retrospective study of the period from July 1988 to July 1994. Of the 415 patients (12.6 percent of the total cases studied) who were positive by the serum test, only 249 (7.6 percent of the total cases studied) were also positive when cerebrospinal fluid was assayed. Thus, 40 percent of the positives (5.6 percent of the total) were identifiable by analysis of serum but not of cerebrospinal fluid. Neisseria meningitidis accounted for 77.7 percent (129) and Haemophilus influenzae for 22.3 percent (37) of the positive results obtained only when serum was examined. These...

Bacterial antigen detection in cerebrospinal fluid by the latex agglutination test.

Eighty purulent cerebrospinal fluid (CSF) samples from patients with clinical evidence of meningitis were studied using the Directigen latex agglutination (LA) kit to determine the presence of bacterial antigen in CSF. The results showed a better diagnostic performance of the LA test than bacterioscopy by Gram stain, culture and counterimmunoelectrophoresis (CIE), as far as Neisseria meningitidis groups B and C, and Haemophilus influenzae type b are concerned, and a better performance than bacterioscopy and culture considering Streptococcus pneumoniae. Comparison of the results with those of culture showed that the LA test had the highest sensitivity for the Neisseria meningitidis group C. Comparing the results with those of CIE, the highest levels of sensitivity were detected for N. meningitidis groups B and C. Regarding specificity, fair values were obtained for all organisms tested. The degree of K agreement when the LA test was compared with CIE exhibited better K indices of agreement for N. meningitidis groups B and C.

Bacterial agents isolated from cerebrospinal fluid of patients with acquired immunodeficiency syndrome (AIDS) and neurological complications.

Cerebrospinal fluid (CSF) samples from 2083 patients with acquired immunodeficiency syndrome (AIDS) and neurological complications were bacteriologically examined during a period of 7 years (1984-1990). The percentage of patients who had at least one bacterial agent cultured from the CSF was 6.2%. Mycobacterium tuberculosis was the most frequently isolated agent (4.3%), followed by Mycobacterium avium complex or MAC (0.7%), Pseudomonas spp (0.5%), Enterobacter spp (0.4%), and Staphylococcus aureus (0.3%). Among 130 culture positive patients, 89 (68.5%) had M. tuberculosis and 15 (11.6%) had MAC. The frequency of bacterial isolations increased from 1988 (5.2%) to 1990 (7.2%), partly due to the increase in MAC isolations. Bacterial agents were more frequently isolated from patients in the age group 21-30 years and from women (p < 0.05).

Production and immunochemical characterization of Neisseria meningitidis group B antiserum for the diagnosis of purulent meningitis.

Unlike Neisseria meningitidis groups A, C, Y and W135, the group B capsular polysaccharide has been shown to be chemically and immunologically identical to the capsular polysaccharide of Escherichia coli K1. Both components are sialic acid homopolymers and are poorly immunogenic. Nevertheless, due to the high incidence of Neisseria meningitidis group B meningitis in the population of the State of São Paulo, preparing antiserum to this serogroup for diagnostic purposes has become a matter of high priority. Of the many immunization schemes proposed, intravenous inoculation of whole bacteria previously inactivated with formaldehyde and simultaneous intradermal inoculation with a mixture of the bacterial polysaccharide fraction and whole bacteria in complete Freund;s adjuvant have produced the best results. The antiserum was treated with immunoadsorbents prepared with aluminum chloride and protein and/or polysaccharide antigens from each of the following heterologous bacteria: Haemophilus influenzae type b, Streptococcus pneumoniae, Escherichia coli other than K1, and Staphylococcus aureus, in order to eliminate cross-reactivity. For quality control analysis, the antiserum was assessed by the immunodiffusion, counterimmunoelectrophoresis, dot-ELISA, and immuno-blot techniques against homologous antigens. Specificity was obtained after treating the antiserum with Haemophilus influenzae type b polysaccharide immunosorbent.

Production and immunochemical characterization of Neisseria meningitidis group B antiserum for the diagnosis of purulent meningitis

Unlike Neisseria meningitidis groups A, C, Y and W135, the group B capsular polysaccharide has been shown to be chemically and immunologically identical to the capsular polysaccharide of Escherichia coli K1. Both components are sialic acid homopolymers and are poorly immunogenic. Nevertheless, due to the high incidence of Neisseria meningitidis group B meningitis in the population of the State of São Paulo, preparing antiserum to this serogroup for diagnostic purposes has become a matter of high priority. Of the many immunization schemes proposed, intravenous inoculation of whole bacteria previously inactivated with formaldehyde and simultaneous intradermal inoculation with a mixture of the bacterial polysaccharide fraction and whole bacteria in complete Freund;s adjuvant have produced the best results. The antiserum was treated with immunoadsorbents prepared with aluminum chloride and protein and/or polysaccharide antigens from each of the following heterologous bacteria: Haemophilus influenzae type b, Streptococcus pneumoniae, Escherichia coli other than K1, and Staphylococcus aureus, in order to eliminate cross-reactivity. For quality control analysis, the antiserum was assessed by the immunodiffusion, counterimmunoelectrophoresis, dot-ELISA, and immuno-blot techniques against homologous antigens. Specificity was obtained after treating the antiserum with Haemophilus influenzae type b polysaccharide immunosorbent.

Biotypes and serotypes of Haemophilus influenzae from patients with meningitis in the city of São Paulo, Brazil.

A total of 1,094 Haemophilus influenzae isolates from cerebrospinal fluid were examined by biochemical and serological means. Most of them belonged to biotype I (70.9%) and to serotype b (99.4%). The relationship of biotypes I and II to the ages of the patients was shown to be significant (P < 0.001).

Protective efficacy of a serogroup B meningococcal vaccine in Sao Paulo, Brazil.

Serogroup B Neisseria meningitidis is the most common cause of epidemic meningococcal disease in developed countries. Until recently no vaccine has been available for prevention of infection with this organism. In an attempt to control epidemic serogroup B meningococcal disease in greater Sao Paulo, Brazil, during 1989 and 1990, a Cuban-produced outer-membrane-protein-based serogroup B meningococcal vaccine was given to about 2.4 million children aged from 3 months to 6 years. We have done a case-control study to estimate the efficacy of the vaccine in greater Sao Paulo. Microbiologically confirmed cases of serogroup B meningococcal disease were identified through hospital-based surveillance. Controls were matched by neighbourhood and age. Vaccination status was confirmed by inspection of vaccination cards. Between June, 1990, and June, 1991, 112 patients and 409 matched controls with confirmed vaccine status were enrolled. Estimated vaccine efficacy varied by age: 48 months or older = 74% (95% Cl 16 to 92%), 24 to 47 months = 47% (-72 to 84%), and less than 24 months = -37% (< -100 to 73%). Our results suggest that the Cuban-produced vaccine may be effective for prevention of serogroup B meningococcal disease in older children and adults.

PIP: In 1990, researchers compared data on 112 3 month-6 year old children who received a Cuban produced, outer-membrane-protein-based serogroup B meningococcal vaccine (cases) and lived in greater Sao Paulo, Brazil with data on 409 age and neighborhood matched controls to determine the protective efficacy of the vaccine against serogroup B meningococcal disease (Neisseria meningitidis). Health workers began administering the vaccine in 1989 to control an epidemic of serogroup B meningococcal disease in the area. In fact, in mid-1989 and early 1990, the rates of serogroup B meningococcal disease in 1-6 year old children in Sao Paulo were 2.07/100,000 and 2.3/100,000, respectively. Even though only 44% of serogroup B meningococcal isolates corresponded with the vaccine type strain (B:4:P1:15), many isolates had man of the same serotype or subtype antigens as the vaccine type strain. Thus the vaccine was able to protect against some other serogroup B meningococcal strains other than the vaccine type strain. Vaccine efficacy for 4-year old children was 74%, but was much lower for 24-47 month old children (47%) and 24-month old children (-37%). The change in the log odds ratio for vaccination by age was linear and significant (p=.057). The researchers suggested that poor vaccine efficacy among younger children may reflect a need for more boosting to achieve protective levels of immunity. The results showed that the Cuban-produced vaccine could contribute to control of outbreaks of serogroup B meningococcal disease by protecting older children and adults from the disease. Researchers need to conduct additional studies of the vaccine and other possible serogroup B meningococcal vaccines.

Sorotipos de Streptococcus pneumoniae isolados de líquido cefalorraquidiano no período de 1977-1988 na cidade de Säo Paulo, Brasil / Serotypes of Streptococcus pneumoniae isolated from cerebrospinal fluid in 1977-1988 in Säo Paulo city, Brasil

Desde 1977, o Instituto Adolfo Lutz (IAL) vem promovendo a sorotipagem do S. pneumoniae ou pneumococo de infecçöes causadas por esta bactéria. As cepas isoladas têm sido encoaminhadas ao WHO Pneumococcal Reference Center, Pensilvania, E.U.A.. De 1977 a 1988, 1.000 cepas de pneumococo isoladas de LCR foram sorotipadas, de acordo com a nomenclatura dinamarquesa, e 60 sorotipos foram sorotipadas, de acordo com a nomenclatura dinamarquesa, e 60 sorotipos foram identificados. A maior freqüência foi do sorotipo 1, secundado por 6B, 18C, 14, 5, 3, 6A, 23F, 19F e 38. Estes sorotipos distribuídos segundo faixas etárias demonstraram incidência variável, notando-se uma certa peculiaridade, ou seja, a predominância do sorotipo 3 no grupo acima de 50 anos. Nos 12 anos considerados, 25 sorotipos apresentaram uma certa uniformidade na freqüência e o mesmo foi observado com relaçäo às estaçöes climáticas, apenas com um número maior de infecçöes meníngeas nos meses mais frios. Considerando a gravidade das infecçöes pneumocócicas notadamente as meningites, e a pouca informaçäo relativa aos sorotipos pneumocócicos que ocorrem na regiäo, julgamos importante essa informaçäo relativa aos sorotipos, uma vez que tem sido usadas, com sucesso, vacinas polissacarídicas na prevençäo dessas infecçöes

[Serotypes of Streptococcus pneumoniae isolated from cerebrospinal fluid in 1977-1988 in São Paulo City, Brazil]. / Sorotipos de Streptococcus pneumoniae isolados de líquido cefalorraquidiano no período de 1977-1988 na cidade de São Paulo, Brasil.

Since 1977, the Instituto Adolfo Lutz (IAL) is having interest in the serotyping of S. pneumoniae or pneumococcus from infections caused by this bacteria. The isolated strains have been sent to the WHO Pneumococcal Reference Center, Pennsylvania, U.S.A.. From 1977 to 1988, 1.000 pneumococcus strains isolated from cerebrospinal fluid were typed, according to Danish nomenclature, and 60 serotypes were identified. The most frequent serotypes were 1, 6B, 18C, 14, 5, 3, 6A, 23F, 19F, and 38. Among different age groups, they showed a variable incidence, with serotype 6B in the ages of zero to almost 2 years old, serotype 1 in the age group of 2 to 50 years old, and serotype 3 in the ages over 50. During the 12 years study, 25 serotypes showed some uniformity in the frequency, the same as with the seasonal fluctuations. Concerning the severity of the pneumococcal infections, chiefly meningitis, and the few information related to pneumococcus serotypes which occur in the area, it was considered of high relevance to have the information of serotypes, once polysaccharide vaccines have been employed with success to prevent these infections.

Febre purpúrica brasileira. Caracterizaçäo rápida das cepas invasoras de Haemophilus aegyptius / Brazilian purpuric fever. Fast characterization of invasive strains of Haemophilus aegyptius

Cepas de H. aegyptus isoladas em surtos de Febre Purpúrica Brasileira (FPB) no Brasil, foram caracterizadas pelo método de aglutinaçäo em lâmina utilizando um anti-soro produzido com cepa de H. aegyptius isolada de cultura de sangue de paciente com FPB. Através desse método foi possível identificar cepas de H. aegyptius responsáveis por surtos de conjuntivite com características antigênicas iguais às cepas isoladas de FPB. A sensibilidade e especificidade da soroaglutinaçäo em lâmina foi de 97,7% e 89,6% respectivamente, podendo ser utilizado como método de triagem em estudos de conjuntivites purulentas, para detectar cepas invasivas de H. aegyptius associada a FPB, possibilitando assim a implantaçäo de medias que ampliem a eficiência na prevençäo e na vigilância epidemiológica da doença