Phenotyping and genotyping methods applied to investigate the relatedness of Brazilian isolates of Enterobacter cloacae

In order to evaluate the resolving power of several typing methods to identify relatedness among Brazilian strains of Enterobacter cloacae, we selected twenty isolates from different patients on three wards of a University Hospital (Orthopedics, Nephrology, and Hematology). Traditional phenotyping methods applied to isolates included biotyping, antibiotic sensitivity, phage-typing, and O-serotyping. Plasmid profile analysis, ribotyping, and macrorestriction analysis by pulsed-field gel electrophoresis (PFGE) were used as genotyping methods. Sero- and phage-typing were not useful since the majority of isolates could not be subtyped by these methods. Biotyping, antibiogram and plasmid profile permitted us to classify the samples into different groups depending on the method used, and consequently were not reliable. Ribotyping and PFGE were significantly correlated with the clinical epidemiological analysis. PFGE did not type strains containing nonspecific DNase. Ribotyping was the most discriminative method for typing Brazilian isolates of E. cloacae

Phenotyping and genotyping methods applied to investigate the relatedness of Brazilian isolates of Enterobacter cloacae.

In order to evaluate the resolving power of several typing methods to identify relatedness among Brazilian strains of Enterobacter cloacae, we selected twenty isolates from different patients on three wards of a University Hospital (Orthopedics, Nephrology, and Hematology). Traditional phenotyping methods applied to isolates included biotyping, antibiotic sensitivity, phage-typing, and O-serotyping. Plasmid profile analysis, ribotyping, and macrorestriction analysis by pulsed-field gel electrophoresis (PFGE) were used as genotyping methods. Sero- and phage-typing were not useful since the majority of isolates could not be subtyped by these methods. Biotyping, antibiogram and plasmid profile permitted us to classify the samples into different groups depending on the method used, and consequently were not reliable. Ribotyping and PFGE were significantly correlated with the clinical epidemiological analysis. PFGE did not type strains containing nonspecific DNase. Ribotyping was the most discriminative method for typing Brazilian isolates of E. cloacae.

Evaluation of the Antimicrobial Activity of Sparfloxacin, Relative to Other Oral Antibiotics Against 1,125 Bacterial Isolates from 10 Medical Centers in Brazil.

A multicenter study was carried out in order to compare the in vitro activity of sparfloxacin to ciprofloxacin, amoxicillin/clavulanic acid, cephalexin, cefuroxine and azithromycin, against 1,125 microorganisms recently isolated from clinical specimens, most of them representative of respiratory tract infections. Sparfloxacin demonstrated potent action and was more active than the beta-lactam agents and azithromycin against most of the bacterial strains tested. Sparfloxacin was more potent (96% and 95% sensitivity, with MIC(90) of 0.19µg/mL and 0.5µg/mL, respectively)than the other antimicrobial agents tested against the Enterobacteriaceae family (Escherichia coli and Elebsiella pneumoniae). It was found to be equivalent in activity to ciprofloxacin (96% and 91% sensitivity and MIC(90) of 0.25 and 0.75µg/mL, respectively). Sparfloxacin was also found to be very active against the most fastidious microorganisms commonly associated to respiratory tract infections such as the penicillin-susceptible and resistant Streptococcus pneumoniae (MIC(90) 0.5µg/mL and 0.38µg/mL, respectively), ampicillin-susceptible and -resistant Haemophilus influenzae (MIC(90) 0.016µg/mL and 0.38µg/mL, respectively), beta-lactamase producing Moraxella catarrhalis (MIC(90) 0.032µg/mL) and non beta-lactamase producing M.catarrhalis (MIC(90) 0.5µg/mL).

Chagas' disease: an algorithm for donor screening and positive donor counseling

Classical serological screening assays for Chagas' disease are time consuming and subjective. The objective of the present work is to evaluate the enzyme immuno-assay (ELISA) methodology and to propose an algorithm for blood banks to be applied to Chagas' disease. Seven thousand, nine hundred and ninety nine blood donor samples were screened by both reverse passive hemagglutination (RPHA) and indirect immunofluorescence assay (IFA). Samples reactive on RPHA and/or IFA were submitted to supplementary RPHA, IFA and complement fixation (CFA) tests. This strategy allowed us to create a panel of 60 samples to evaluate the ELISA methodology from 3 different manufacturers. The sensitivity of the screening by IFA and the 3 different ELISA's was 100%. The specificity was better on ELISA methodology. For Chagas disease, ELISA seems to be the best test for blood donor screening, because it showed high sensitivity and specificity, it is not subjective and can be automated. Therefore, it was possible to propose an algorithm to screen samples and confirm donor results at the blood bank.

Os testes sorológicos clássicos utilizados na triagem de doadores de sangue são trabalhosos e subjetivos. O objetivo do presente trabalho é o de avaliar a metodologia imuno-enzimática (ELISA) e propor um algorítmo para doença de Chagas em bancos de sangue. Foram estudados 7999 doadores de sangue e/ou componentes cujas amostras foram testadas com o objetivo de tria-las sorologicamente para doença de Chagas utilizando hemaglutinação passiva reversa (RPHA) e imunofluorescência indireta (IFA). As amostras reativas em pelo menos uma destas metodologias, foram retestadas com reativos diferentes por RPHA, IFA e fixação de complemento (CFA). Esta estratégia nos permitiu criar um painel de 60 amostras com as quais tornou-se possível a avaliação do método imunoenzimático (ELISA). A sensibilidade da triagem dos doadores pelos métodos ELISA e IFA foi de 100%. A especificidade foi melhor para a metodologia imunoenzimática. O teste ELISA parece ser o ideal para triagem em bancos de sangue pois é altamente sensível, específico, não é subjetivo e pode ser automatizado. Desta forma, torna-se possível a formulação de um algorítimo a ser utilizado na triagem sorológica e confirmação de resultados em doadores de bancos de sangue.

Chagas' disease: an algorithm for donor screening and positive donor counseling.

Classical serological screening assays for Chagas' disease are time consuming and subjective. The objective of the present work is to evaluate the enzyme immuno-assay (ELISA) methodology and to propose an algorithm for blood banks to be applied to Chagas' disease. Seven thousand, nine hundred and ninety nine blood donor samples were screened by both reverse passive hemagglutination (RPHA) and indirect immunofluorescence assay (IFA). Samples reactive on RPHA and/or IFA were submitted to supplementary RPHA, IFA and complement fixation (CFA) tests. This strategy allowed us to create a panel of 60 samples to evaluate the ELISA methodology from 3 different manufacturers. The sensitivity of the screening by IFA and the 3 different ELISA's was 100%. The specificity was better on ELISA methodology. For Chagas disease, ELISA seems to be the best test for blood donor screening, because it showed high sensitivity and specificity, it is not subjective and can be automated. Therefore, it was possible to propose an algorithm to screen samples and confirm donor results at the blood bank.

Raiva humana tratada com interferon por via intratecal / Human rabies treated with interferon by intramuscular way

Relata-se um caso de raiva humana submetido a tratamento com interferon leucocitário humano injetado por via ventricular, através de reservatório de Rickham, e parte por via intramuscular. As doses diárias injetadas foram de 5 x10**6 unidades por via intraventricular e de 20 x 10**6 unidades por via intramuscular, a partir do 7§ dia de doença durante nove dias, quando entäo ocorreu o óbito em conseqüência de sepse por Klebsiella pneumoniae (16§ dia de doença). O vírus da raiva foi isolado inicialmente através de biopsia do tecido nervoso cerebral. O nível de interferon no sangue e no líquor era inferior a 10 U/ml antes do início do tratamento. O esquema terapêutico utilizado propiciou títulos elevados de interferon no sangue e no líquor 10 a 2.000 vezes maiores que aqueles atingidos usualmente na infecçäo natural. O nível residual, após 24 horas, foi de 212 ñ 14 U no sangue e 451 ñ 81 U no líquor. Foi observada menor soroconversäo (anticorpos neutralizantes) contra o vírus rábico (1/75 a partir do 13§ dia), em conseqüência da imunossupressäo induzida pelo interferon, provavelmente

Seroepidemiological studies of HIV-1 infection in large Brazilian cities.

The prevalence of HIV-1 antibodies in selected groups of individuals from Rio de Janeiro, São Paulo and Santos was determined retrospectively. These groups and respective prevalences were: hemophiliac patients from Rio de Janeiro (1983-1984) 98.0%; polytransfused hemodialysis patients from São Paulo (1985-1986) 3.0% and (1987) 7.7%; intravenous drug addicts from São Paulo and Rio de Janeiro (1986-1987) 15.9%; male prisoners from São Paulo (1988) 12.5%, and pregnant women from Santos (1988-1989) 3.6%. These data stress the magnitude of AIDS in Brazil.

Legionelose no Brasil: inquerito sorologico entre doadores de sangue e trabalhadores em unidades de terapia intensiva de tres hospitais de Sao Paulo. / Legionellosis in Brazil: serological survey among blood donors and personnel of intensive care units of 3 hospitals of Sao Paulo

Com a finalidade de constatar a presenca do agente causal da legionelose no Brasil, foi realizado um inquerito sorologico pela tecnica de imunofluorescencia indireta em quatro grupos de individuos que vivem e trabalham na cidade de S. Paulo. Tres grupos foram constituidos por funcionarios de tres Unidades de Terapia Intensiva de tres hospitais gerais da cidade, e um quarto grupo de doadores de sangue de um Banco de Sangue (120 amostras de sangue), o teste resultou positivo em apenas 2.5%. Estes resultados sugerem que a legionelose-infeccao e, provavelmente, mais prevalente em pessoal que trabalha em ambientes onde se acredita que a bacteria seja mais prevalente, como, por exemplo, em Unidades de Terapia Respiratoria Intensiva

Naturally acquired antibodies to tetanus toxin in humans and animals from the galápagos islands.

A serologic survey using a highly sensitive enzyme-linked immunosorbent assay confirmed the anticipated finding of naturally acquired antibodies to tetanus toxin both in humans and animals on the Galápagos Islands. In 57 inhabitants (mean age, 31.3 years) who had not been vaccinated against tetanus, antibody to tetanus toxin was detected in the blood in varying titers (geometric mean [reciprocal] titer [GMT], 0.015 international units [IU]/ml). In one individual the titer of antibody was greater than 12.5 IU/ml. Two individuals who had never been vaccinated against tetanus but who had reported having had clinical tetanus had titers of antibody to tetanus toxin of 0.02 IU/ml and 0.3 IU/ml, respectively. All nine of the animals studied showed antibody to tetanus toxin (GMT, 0.028 IU/ml).

Tratamento especifico do tetano com uma fracao - F(ab') - da imunoglobulina antitetanica injetada por via raquidiana. / Specific treatment of tetanus with a fraction - F(ab') - of the antitetanic immunoglobulin injected intrathecally

Sao apresentados os resultados preliminares do tratamento especifico de 26 casos de tetano humano com a fracao F(ab') da antitoxina tetanica equina injetada por via intratecal. O indice de letalidade observado foi de 3,8%. Considerando-se apenas os casos de maior gravidade (grau III) o indice de letalidade foi de 10%. Tais resultados encorajadores vem corroborar os resultados de experimentacoes previas em animais de laboratorio, realizados por outros pesquisadores, onde a importancia da via sub-aracnoidea e de fracoes de baixo peso molecular de gamaglobulina antitetanica foram demonstradas importantes para permitir o acesso da antitoxina aos neuronios das membranas pre-sinapticas da medula espinal a fim de deslocar e neutralizar a toxina fixada aos receptores dos gangliosideos

Tratamento do herpes genital e labial com interferon. / Treatment of genital and labial herpes with interferon

Foram analisadas as caracteristicas dos herpesvirus e os mecanismos imunologicos implicados na patogenese da doenca herpetica. Os fundamentos que explicam as perspectivas promissoras do tratamento da doenca herpetica com interferon foram analisados em funcao dos atuais conhecimentos imunologicos virulogicos e patogenicos sobre a doenca. Foi possivel isolar o herpesvirus em 50% dos especimes enviados ao laboratorio (nao foi feita a tipagem dos virus). Dentre os parametros clinicos utilizados, houve desaparecimento da dor e prurido em 89% dos pacientes e da queimacao em todos. Nao houve qualquer insucesso do tratamento, ja que 50% estavam totalmente curados e os restantes em vias de cura (crostas) no 4o. dia apos o inicio da terapeutica local com pomada de interferon