Relação genética entre cepas de Neisseria meningitidis sorogrupo B sorotipo 4

We compared the results obtained by serotyping of PorB epitopes using an expanded panel of monoclonal antibodies (mAb) including mAb 7 and mAb 10, with results obtained by RFLP of rRNA genes (ribotyping). The purpose of this study was to assess the correlation between phenotypic- and genotypic- methods for typing N. meningitidis. The ribotypes obtained using ClaI or EcoRV endonucleases grouped the strains in seven and two different patterns, respectively. This additional characterization of PorB epitopes improved the correlation between these two methods of typing N. meningitidis.Nós comparamos os resultados obtidos pela sorotipagem usando um painel de anticorpos monoclonais (mAb) ao qual se incluíam os mAbs 7 e 10, com os resultados obtidos pelo RFLP do gene rRNA (ribotipagem). O propósito deste estudo foi avaliar a correlação entre os métodos fenotípico e genotípico de tipagem de N. meningitidis. Os ribotipos obtidos usando as endonucleases de restrição ClaI e EcoRV foram hábeis em dividir as cepas em sete e dois diferentes perfis de restrição, respectivamente. A caracterização adicional de epítopos na proteína PorB melhorou a correlação entre os dois métodos de tipagem de N. meningitidis

The use of oligonucleotide probes for meningococcal serotype characterization

In the present study we examine the potential use of oligonucleotide probes to characterize Neisseria meningitidis serotypes without the use of monoclonal antibodies (MAbs). Antigenic diversity on PorB protein forms the bases of serotyping method. However, the current panel of MAbs underestimated, by at least 50% the PorB variability, presumably because reagents for several PorB variable regions (VRs) are lacking, or because a number of VR variants are not recognized by serotype-defining MAbs12. We analyzed the use of oligonucleotide probes to characterize serotype 10 and serotype 19 of N. meningitidis. The porB gene sequence for the prototype strain of serotype 10 was determined, aligned with 7 other porB sequences from different serotypes, and analysis of individual VRs were performed. The results of DNA probes 21U (VR1-A) and 615U (VR3-B) used against 72 N. meningitidis strains confirm that VR1 type A and VR3 type B encode epitopes for serotype-defined MAbs 19 and 10, respectively. The use of probes for characterizing serotypes possible can type 100% of the PorB VR diversity. It is a simple and rapid method specially useful for analysis of large number of samples

Meningococcal Disease Caused by Neisseria meningitidis Serogroup B Serotype 4 in São Paulo, Brazil, 1990 to 1996

A large epidemic of serogroup B meningococcal disease (MD), has been occurring in greater São Paulo, Brazil, since 1988.21 A Cuban-produced vaccine, based on outer-membrane-protein (OMP) from serogroup B: serotype 4: serosubtype P1.15 (B:4:P1.15) Neisseria meningitidis, was given to about 2.4 million children aged from 3 months to 6 years during 1989 and 1990. The administration of vaccine had little or no measurable effects on this outbreak. In order to detect clonal changes that could explain the continued increase in the incidence of disease after the vaccination, we serotyped isolates recovered between 1990 and 1996 from 834 patients with systemic disease. Strains B:4:P1.15, which was detected in the area as early as 1977, has been the most prevalent phenotype since 1988. These strains are still prevalent in the area and were responsible for about 68% of 834 serogroup B cases in the last 7 years. We analyzed 438 (52%) of these strains by restriction fragment length polymorphism (RFLPs) of rRNA genes (ribotyping). The most frequent pattern obtained was referred to as Rb1 (68%). We concluded that the same clone of B:4:P1.15-Rb1 strains was the most prevalent strain and responsible for the continued increase of incidence of serogroup B MD cases in greater São Paulo during the last 7 years in spite of the vaccination trial

Genetic structure of Neisseria meningitidis serogroup C epidemic strains in South Brazil

In the present study we report the results of an analysis, based on serotyping, multilocus enzyme electrophoresis (MEE), and ribotyping of N. meningitidis serogroup C strains isolated from patients with meningococcal disease (MD) in Rio Grande do Sul (RS) and Santa Catarina (SC) States, Brazil, as the Center of Epidemiology Control of Ministry of Health detected an increasing of MD cases due to this serogroup in the last two years (1992-1993). We have demonstrated that the MD due to N.meningitidis serogroup C strains in RS and SC States occurring in the last 4 years were caused mainly by one clone of strains (ET 40), with isolates indistinguishable by serogroup, serotype, subtype and even by ribotyping. One small number of cases that were not due to an ET 40 strains, represent closely related clones that probably are new lineages generated from the ET 40 clone referred as ET 11A complex. We have also analyzed N.meningitidis serogroup C strains isolated in the greater São Paulo in 1976 as representative of the first post epidemic year in that region. The ribotyping method, as well as MEE, could provide useful information about the clonal characteristics of those isolates and also of strains isolated in south Brazil. The strains from 1976 have more similarity with the actual endemic than epidemic strains, by the ribotyping, sulfonamide sensitivity, and MEE results. In conclusion, serotyping with monoclonal antibodies (C:2b:P1.3), MEE (ET 11 and ET 11A complex), and ribotyping by using ClaI restriction enzyme (Rb2), were useful to characterize these epidemic strains of N.meningitidis related to the increased incidence of MD in different States of south Brazil. It is mostly probable that these N.meningitidis serogroup C strains have poor or no genetic corelation with 1971-1975 epidemic serogroup C strains. The genetic similarity of members of the ET 11 and ET 11A complex were confirmed by the ribotyping method by using three restriction endonucleases.No presente estudo, nós reportamos os resultados de uma análise, baseada na sorotipagem, multilocus enzimático (MEE) e ribotipagem de N. meningitidis sorogrupo C isoladas de paciente com doença meningocócica no Rio Grande do Sul (RS) e Santa Catarina (SC), onde o Centro de Controle Epidemiológico do Ministério da Saúde detectou um aumento do número de casos de doença meningocócica (DM) devido a este sorogrupo nos últimos 2 anos (1992-1993). Nós demonstramos que a DM devido a cepas de N. meningitidis sorogrupo C no RS e SC que ocorreram nos últimos 4 anos foi devido principalmente por um clone (ET 40), com isolados indistinguíveis por sorogrupo, sorotipo, subtipo e até por ribotipagem. Um pequeno número de casos que não foram devidos a cepas do ET 40 representaram um grupo geneticamente relacionado, que provavelmente é uma nova linhagem gerada do clone ET 40, referido como complexo ET 11 A. Nós também analisamos cepas de N. meningitidis sorogrupo C isoladas na grande São Paulo em 1976 como um grupo representativo do primeiro ano pós-epidêmico na região. A ribotipagem, bem como MEE, puderam fornecer informações sobre as características clonais das cepas isoladas no período pós-epidêmico e também no Sul do Brasil. As cepas de 1976 possuem mais similaridades com as cepas endêmicas atuais do que com as cepas epidêmicas (1992-1993) por ribotipagem, sensibilidade a sulfonamida e MEE. Em conclusão, sorotipagem com anticorpos monoclonais (C:2b:P1.3), MEE (complexo ET11 e ET11A) e ribotipagem usando a enzima de restrição ClaI, foram úteis em caracterizar estas cepas epidêmicas de N. meningitidis relacionadas com o aumento da incidência da DM em diferentes estados do sul do Brasil. É muito provável que estas cepas de N. meningitidis sorogrupo C possuam pouca ou nenhuma correlação genética com as cepas epidêmicas sorogrupo C de 1971-1975. A similaridade genética dos membros do complexo ET 11 e ET 11A foram confirmadas por ribotipagem usando-se 3 enzimas de restrição

Listeria monocytogenes in renal transplant recipients Listeria monocytogenes em pacientes pós-transplante renal

Five cases of Listeria monocytogenes bacteriemia were observed from April to December 1985, among renal transplant recipients from the same hospital in São Paulo, Brazil. The patients were adults (mean age: 40.6 years), and the basic complain was fever, with no report of meningeal syndrome. Laboratory tests revealed the presence of two serovars, 1/2a and 4b, which were classified into three lysotypes. The four strains of serovar 4b showed the same antibiotype, with resistance to cefoxitin, clindamycin, oxacillin and penicillin.<br>No período de abril a dezembro de 1985, foram observados cinco casos de listeriose em transplantados renais num mesmo hospital de São Paulo, SP. Os pacientes eram adultos (média de 40,6 anos) tendo como queixa básica a febre. Laboratorialmente, em todos foram reconhecidos Listeria monocytogenes, caracterizada por dois sorovares 1/2a e 4b e três lisotipos distintos. As amostras do sorovar 4b apresentaram o mesmo antibiotipo: resistentes à cefoxitina, clindamicina, oxacilina e penicilina