Frequência laboratorial e resistência antimicrobiana de enteropatógenos bacterianso isolados em crianças na primeira infância na região do ABC Paulista, 2015 - 2016 / Laboratory frequency and antimicrobial resistance of the enteropathogens bacteria isolated from infants at early childhood in the region of ABC Paulista, 2015 - 2016

O objetivo deste estudo foi avaliar o perfil dos enteropatógenos bacterianos isolados em crianças menores de 5 anos durante casos de diarreia em instituições de 4 municípios do Estado de São Paulo, durante 2015 e 2016. A coleta das fezes foi realizada em 107 crianças, 78 (72,9%) crianças com diarreia e 29 (27,1%) crianças sem diarreia. A metodologia foi coprocultura, identificação bacteriana e teste de sensibilidade aos antimicrobianos. Quarenta e seis das 107 (43%) amostras clínicas apresentaram crescimento de enteropatógenos. Amostras de Escherichia coli enteropatogênicas (EPEC), Escherichia coli enteroagregativas (EAEC) e Salmonella enterica subsp houtenae foram as mais frequentemente isoladas entre as crianças. Do total de crianças estudadas, três delas apresentaram co-infecção por 2 agentes etiológicos diferentes: EPEC/EAEC e Salmonella enterica subsp houtenae/EAEC. A maior ocorrência entre os 49 agentes etiológicos isolados foi EPEC (24/49, 49%), seguido de EAEC (14/49, 28,6%). Duas amostras de EPEC pertencentes ao sorotipo O109:H21 foram sensíveis aos antimicrobianos testados, enquanto outras duas pertencentes ao sorotipo O156:H1 foram resistentes a gentamicina e a amicacina e estreptomicina, respectivamente. Duas amostras de EAEC pertencentes ao mesmo sorotipo O80:H10 e duas EAEC O15:H2 apresentaram multirresistência, pelo menos, ao ácido nalidíxico, sulfametoxazol e tetraciclina. Podemos sugerir que crianças frequentadoras de três instituições diferentes, que apresentaram agregados de casos de diarreia sugestivo de surto, eram portadoras de clones bacterianos de amostras de EPEC ou EAEC, por pertencerem ao mesmo sorotipo e com semelhante perfil de sensibilidade. Nossos resultados são preocupantes e mostram que a vigilância epidemiológica antimicrobiana constante deve ser garantida para o monitoramento do surgimento de clones resistentes e para estabelecer estratégias para a prevenção e c ontrole de surtos e epidemias

This study aimed at evaluating the profile of enteropathogens isolated from children under 5 years of age, during the occurrence of cases of diarrhe a in the institutions of four municipalities in the State of São Paulo in 2015 and 2016. Feces samples were collected from 107 children, 78 (72.9%) with diarrhea and 29 (27.1%) without. The employed methodologies were copro-culture, bacterial identification and antimicrobial susceptibility testing. Forty-six (46%) of 107 clinical samples presented growth of enteropathogens. Enteropathogenic Escherichia coli (EPEC), enteroaggregative Escherichia coli (EAEC) and Salmonella enterica subsp houtenae were the mostly frequent isolated from children. Of the total number of the studied children, three of them presented co-infection with two etiological agents: EPEC/EAEC and Salmonella enterica subsp houtenae/EAEC. The highest occurrence among the isolated etiologic agents was EPEC (24/49, 49%), followed by EAEC (14/49, 28.6%). Two EPEC strains belonged to the O109:H21 serotype were sensitive to the tested antimicrobials, whereas two belonging to the O156:H1 serotype were resistant to gentamicin and amicacin and streptomycin, respectively. Two EAEC strains of the same serotype O80:H10 and two EAEC O15:H2 presented multi-resistance at least to nalidixic acid, sulfamethoxazole and tetracycline. It may suggest that the children attending three different institutions, who had clusters of cases of diarrhoea carried the bacterial clones of EPEC or EAEC strains, because they belonged to the same serotype and show a similar sensitivity profile. The results found in the present study are worrying and they show that the constant antimicrobial epidemiological surveillance should be ensured for monitoring the emergence of resistant clones, and for establishing strategies for preventing and controlling the outbreaks and epidemics.

Surto de doença trasmitida por alimentos nos municipios de Mauá e Ribeirão Pires - SP / Outbreak of foodborne diseases in the municipalities of Maua and Ribeirao Pires - SP

Doenças de transmissão hídrica e alimentar (DTHA) acarretam importantes problemas econômicos e de saúde pública no mundo atual. Este estudo relata um surto de Doença Transmitida por Alimento - DTA que envolveu 12 pessoas de duas residências localizadas na Região do ABC paulista em dezembro de 2012. Quatro pessoas de uma residência tiveram sintomas de diarreia, cólica abdominal, náusea, vômito, febre e prostração, sendo que apenas duas consumiram o bolo preparado em Ribeirão Pires, SP - Brasil. Outras oito pessoas consumiram o mesmo alimento no município de Mauá e, além dos sintomas citados, houve também registro de insuficiência renal e parada cardiorrespiratória. Dentre os envolvidos, uma menina de oito anos veio a óbito após convulsão e bronco-aspiração. O período variou entre 2 e 22 horas após o consumo do alimento. A amostra de bolo foi analisada segundo a metodologia preconizada pelo BAM-FDA e teve como resultados: Coliformes termotolerantes (NMP = 4,6x104/g); Bacillus cereus (1,5x105 U.F.C./g) e presença de Salmonella Enteritidis em 25 gramas. Clostridium perfringens, Staphylococcus aureus e Listeria monocytogenes não foram isolados. Foram realizadas duas coproculturas que apresentaram resultados positivos para Salmonella Enteritidis. As cepas de Salmonella spp isoladas, tanto no alimento como nas fezes dos pacientes, apresentaram similaridade genética e mesmo perfil de suscetibilidade aos antimicrobianos. Assim, foi constatado o envolvimento do bolo como veiculador de patógenos e ressaltada a importância do trabalho em conjunto das vigilâncias sanitárias e epidemiológicas de ambos os municípios e o laboratório de referência em saúde pública, fundamental na elucidação deste surto.

Surto de doença transmitida por alimentos nos municípios de Mauá e Ribeirão Pires - SP / Outbreak of food-borne disease in the municipalities of Maua´and Ribeirão Pires - SP

Doenças de transmissão hídrica e alimentar (DTHA) acarretam importantes problemas econômicos e de saúde pública no mundo atual. Este estudo relata um surto de Doença Transmitida por Alimento - DTA que envolveu 12 pessoas de duas residências localizadas na Região do ABC paulista em dezembro de 2012. Quatro pessoas de uma residência tiveram sintomas de diarreia, cólica abdominal, náusea, vômito, febre e prostração, sendo que apenas duas consumiram o bolo preparado em Ribeirão Pires, SP - Brasil. Outras oito pessoas consumiram o mesmo alimento no município de Mauá e, além dos sintomas citados, houve também registro de insuficiência renal e parada cardiorrespiratória. Dentre os envolvidos, uma menina de oito anos veio a óbito após convulsão e bronco-aspiração. O período variou entre 2 e 22 horas após o consumo do alimento. A amostra de bolo foi analisada segundo a metodologia preconizada pelo BAM-FDA e teve como resultados: Coliformes termotolerantes (NMP = 4,6x104 /g); Bacillus cereus (1,5x105 U.F.C./g) e presença de Salmonella Enteritidis em 25 gramas. Clostridium perfringens, Staphylococcus aureus e Listeria monocytogenes não foram isolados. Foram realizadas duas coproculturas que apresentaram resultados positivos para Salmonella Enteritidis. As cepas de Salmonella spp isoladas, tanto no alimento como nas fezes dos pacientes, apresentaram similaridade genética e mesmo perfil de suscetibilidade aos antimicrobianos. Assim, foi constatado o envolvimento do bolo como veiculador de patógenos e ressaltada a importância do trabalho em conjunto das vigilâncias sanitárias e epidemiológicas de ambos os municípios e o laboratório de referência em saúde pública, fundamental na elucidação deste surto. (AU)

Foodborne illness are still important public health problems in the world today. This paper reports an outbreak of food poisoning involving 12 people from two residences located in the ABC Region, São Paulo, Brazil in December 2012. Four people of one residence had symptoms of diarrhea, abdominal cramps, nausea, vomiting, fever and prostration, and only two people had eaten the cake. Eight others had consumed the same food in the Mauá city and beyond the symptoms mentioned above, there was kidney failure and cardiac arrest. Among those involved, an eight-year-old girl has died after seizure and bronchial aspiration. The incubation period ranged from 2 to 22 hours. The cake sample was analyzed according to the methods recommended by the FDA-BAM and had the following results:thermotolerant coliforms (NMP = 4.6 X 104 / g); Bacillus cereus (1.5 X 105 CFU / g) and presence of Salmonella Enteritidis in 25 g. Clostridium perfringens, Staphylococcus aureus and Listeria monocytogenes were not isolated. Two stools samples were tested for Salmonella Enteritidis had been positive for both. Salmonella strains isolated in both food and stools of patients has showed genetic similarity and the same profile of antimicrobial susceptibility. Thus, it was found the involvement of the cake as disseminator of pathogens and emphasized the importance of working together of sanitary and epidemiological surveillance of both cities and public health laboratory, fundamental to elucidation of this outbreak. (AU)

Effective characterization of Salmonella Enteritidis by most probable number (MPN) followed by multiplex polymerase chain reaction (PCR) methods.

Nontyphoidal Salmonella (NTS) is a relevant pathogen involved in gastroenteritis outbreaks worldwide. In this study, we determined the capacity to combine the most probable number (MPN) and multiplex polymerase chain reaction (PCR) methods to characterize the most important Salmonella serotypes in raw sewage. A total of 499 isolates were recovered from 27 raw sewage samples and screened using two previously described multiplex PCR methods. From those, 123 isolates were selected based on PCR banding pattern-identical or similar to Salmonella Enteritidis and Salmonella Typhimurium-and submitted to conventional serotyping. Results showed that both PCR assays correctly serotyped Salmonella Enteritidis, however, they presented ambiguous results for Salmonella Typhimurium identification. These data highlight that MPN and multiplex PCR can be useful methods to describe microbial quality in raw sewage and suggest two new PCR patterns for Salmonella Enteritidis identification.

Population Structure Analysis of Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates from Brazil Reveals Predominance of Clonal Complexes 1, 15, and 79.

The population structure of 71 carbapenem-resistantAcinetobacter baumanniiclinical isolates from several hospitals in Brazil was investigated by ApaI pulsed-field gel electrophoresis,blaOXA-51-like subtyping, and multilocus sequence typing (Institute Pasteur scheme). In addition to the predominance of strains carryingblaOXA-23, we detected the presence ofblaOXA-72andblaOXA-231 We observed a predominance of clonal complex 1 (CC1), CC15, and CC79 and representative strains of the worldwide-disseminated international clone I.

Salmonella Alachua: causative agent of a foodborne disease outbreak

Objectives: The aim of this study is to report the occurrence of the first outbreak of food poisoning caused by Salmonella Alachua in Brazil, as well as the antimicrobial susceptibility and the genetic relatedness of Salmonella Alachua strains isolated from clinical and food samples. Material and methods: To elucidate the outbreak, an epidemiological investigation was carried out, and two samples of common food were tested - mayonnaise salad and galinhada (a traditional Brazilian dish of chicken and rice) - according to the Compendium of methods for the microbiological examination of foods. Five stool samples were tested employing classic methods for the isolation and identification of enterobacteria. Strains of Salmonella were characterized for antibiotic susceptibility according to the Clinical and Laboratory Stan- dards Institute guidelines (2013), and submitted to pulsed-field gel electrophoresis analysis, performed according to the Centers for Disease Control and Prevention PulseNet protocol. Results: A total of 94 people were interviewed after ingesting the food, 66 of whom had become ill. A 60-year old female patient who was hospitalized in a serious condition, developed septic shock and died two days after consuming the food. The presence of Salmonella Alachua was confirmed in all the analyzed stool samples, and in the two types of food. The five strains showed higher than minimum inhibitory concentration values of nalidixic acid (≥256 µg/mL) and reduced ciprofloxacin susceptibility (minimum inhibitory concentration = 0.5 µg/mL). The pulsed-field gel electrophoresis analysis revealed indistinguishable patterns in Salmonella Alachua strains isolated from clinical and food samples. Conclusion: The data presented herein confirm the foodborne disease outbreak. They also allowed for the identification of the source of infection, and suggest that products from poultry are potential reservoirs for this serotype, reinforcing the ...

Salmonella Alachua: causative agent of a foodborne disease outbreak.

OBJECTIVES: The aim of this study is to report the occurrence of the first outbreak of food poisoning caused by Salmonella Alachua in Brazil, as well as the antimicrobial susceptibility and the genetic relatedness of Salmonella Alachua strains isolated from clinical and food samples. MATERIAL AND METHODS: To elucidate the outbreak, an epidemiological investigation was carried out, and two samples of common food were tested--mayonnaise salad and galinhada (a traditional Brazilian dish of chicken and rice)--according to the Compendium of methods for the microbiological examination of foods. Five stool samples were tested employing classic methods for the isolation and identification of enterobacteria. Strains of Salmonella were characterized for antibiotic susceptibility according to the Clinical and Laboratory Standards Institute guidelines (2013), and submitted to pulsed-field gel electrophoresis analysis, performed according to the Centers for Disease Control and Prevention PulseNet protocol. RESULTS: A total of 94 people were interviewed after ingesting the food, 66 of whom had become ill. A 60-year old female patient who was hospitalized in a serious condition, developed septic shock and died two days after consuming the food. The presence of Salmonella Alachua was confirmed in all the analyzed stool samples, and in the two types of food. The five strains showed higher than minimum inhibitory concentration values of nalidixic acid (≥256 µg/mL) and reduced ciprofloxacin susceptibility (minimum inhibitory concentration=0.5 µg/mL). The pulsed-field gel electrophoresis analysis revealed indistinguishable patterns in Salmonella Alachua strains isolated from clinical and food samples. CONCLUSION: The data presented herein confirm the foodborne disease outbreak. They also allowed for the identification of the source of infection, and suggest that products from poultry are potential reservoirs for this serotype, reinforcing the importance of warning consumers about the danger of possible contamination.

Identification of new flagellin-encoding fliC genes in Escherichia coli isolated from domestic animals using RFLP-PCR and sequencing methods / Identificação de novas flagelinas codificadas por fliC em Escherichia coli isoladas de animais domésticos utilizando RFLP-PCR e sequenciamento

Identification of Escherichia coli requires knowledge regarding the prevalent serotypes and virulence factors profiles allows the classification in pathogenic/non-pathogenic. However, some of these bacteria do not express flagellar antigen invitro. In this case the PCR-restriction fragment length polymorphism (RFLP-PCR) and sequencing of the fliC may be suitable for the identification of antigens by replacing the traditional serology. We studied 17 samples of E. coli isolated from animals and presenting antigen H nontypeable (HNT). The H antigens were characterized by PCR-RFLP and sequencing of fliC gene. Three new flagellin genes were identified, for which specific antisera were obtained. The PCR-RFLP was shown to be faster than the serotyping H antigen in E. coli, provided information on some characteristics of these antigens and indicated the presence of new genes fliC.

A identificação da Escherichia coli requer conhecimento sobre os sorotipos e fatores de virulência prevalentes permitindo a classificação em patogênico/não patogênico. No entanto, algumas destas bactérias não expressam o antígeno flagelar in vitro. Neste caso, o PCR-restriction fragment length polymorphism (RFLP-PCR) e o sequenciamento do gene fliC podem ser adequados para a identificação desses antígenos, substituindo a sorologia tradicional. Nesta pesquisa foram estudadas 17 amostras de E. coli isoladas de animais e que apresentavam antígeno H não tipável (HNT). Os antígenos H foram caracterizados por PCR-RFLP e sequenciamento do gene fliC. Três novos genes da flagelina foram identificados, para os quais anti-soros específicos foram obtidos. A técnica PCR-RFLP mostrou-se mais rápida que a sorotipagem do antígeno H em E. coli, fornecendo informações sobre algumas características desses antígenos e indicou a presença de novos genes fliC.

Draft genome of a Brazilian avian-pathogenic Escherichia coli strain and in silico characterization of virulence-related genes.

Avian-pathogenic Escherichia coli (APEC) strains cause extraintestinal diseases in avian species. Here, we present the draft genome of an APEC strain (SCI-07) from Brazil that was isolated from skin lesions (gelatinous edema) on the head and periorbital tissues of a laying hen with swollen head syndrome.

Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFlP and DNA sequencing methods.

Escherichia coli has been isolated frequently, showing flagellar antigens that are not recognized by any of the 53 antisera, provided by the most important reference center of E. coli, The International Escherichia and Klebsiella Center (WHO) of the Statens Serum Institute, Copenhagen, Denmark. The objective of this study was to characterize flagellar antigens of E. coli that express non-typeable H antigens. The methods used were serology, PCR-RFLP and DNA sequencing. This characterization was performed by gene amplification of the fliC (flagellin protein) by polymerase chain reaction in all 53 standards E.coli strains for the H antigens and 20 E. coli strains for which the H antigen was untypeable. The amplicons were digested by restriction enzymes, and different restriction enzyme profiles were observed. Anti-sera were produced in rabbits, for the non-typeable strains, and agglutination tests were carried out. In conclusion,the results showed that although non-typeable and typable H antigens strains had similar flagellar antigens, the two types of strains were distinct in terms of nucleotide sequence, and did not phenotypically react with the standard antiserum, as expected. Thirteen strains had been characterized as likely putative new H antigen using PCR-RFLP techniques, DNA sequencing and/or serology.

Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFlP and DNA sequencing methods

Escherichia coli has been isolated frequently, showing flagellar antigens that are not recognized by any of the 53 antisera, provided by the most important reference center of E. coli, The International Escherichia and Klebsiella Center (WHO) of the Statens Serum Institute, Copenhagen, Denmark. The objective of this study was to characterize flagellar antigens of E. coli that express non-typeable H antigens. The methods used were serology, PCR-RFLP and DNA sequencing. This characterization was performed by gene amplification of the fliC (flagellin protein) by polymerase chain reaction in all 53 standards E.coli strains for the H antigens and 20 E. coli strains for which the H antigen was untypeable. The amplicons were digested by restriction enzymes, and different restriction enzyme profiles were observed. Anti-sera were produced in rabbits, for the non-typeable strains, and agglutination tests were carried out. In conclusion,the results showed that although non-typeable and typable H antigens strains had similar flagellar antigens, the two types of strains were distinct in terms of nucleotide sequence, and did not phenotypically react with the standard antiserum, as expected. Thirteen strains had been characterized as likely putative new H antigen using PCR-RFLP techniques, DNA sequencing and/or serology.

Correlation between API 50 CH and multiplex polymerase chain reaction for the identification of vaginal lactobacilli in isolates

Identification of Lactobacillus sp. strains by phenotypic methods may lead to doubtful results possibly interfering in the reliability of the epidemiological and probiotics studies. Therefore this study aimed to determine the best methodology for the identification of the large diversity of lactobacilli species found in the vagina by comparing two techniques, one based on their biochemical profile and other employing molecular biology. A carbohydrate fermentation test (API 50 CH) was compared with multiplex polymerase chain reaction (PCR) for the identification of species of vaginal lactobacilli from 135 healthy women. The kappa index was used to evaluate agreement between the methods. Using the molecular technique, L. crispatus (32.6 percent), L. jensenii (25 percent) and L. gasseri (20.6 percent) were the most frequent species. However, using the biochemical technique, the most frequent species were: L. acidophilus (34.8 percent), L. crispatus (27.2 percent) and L. fermentum (13 percent). Although L. acidophilus was the most frequent specie found by biochemical tests, no strain of this microorganism was detected by PCR. Agreement between the methods was low for identification of all the most common species. Although rates of L. crispatus detected were similar using both methods (32.6 percent and 27.2 percent), agreement between them was relatively low (kappa = 0.52). Conclusions: Our results confirmed the limitation of the biochemical method and the applicability of a previously published molecular method (Multiplex PCR) for the identification of lactobacilli in the vaginal tract, focusing on further necessity of its improvement for also targeting L. vaginalis and L. iners.

Correlation between api 50 ch and multiplex polymerase chain reaction for the identification of vaginal lactobacilli in isolates.

UNLABELLED: Identification of Lactobacillus sp. strains by phenotypic methods may lead to doubtful results possibly interfering in the reliability of the epidemiological and probiotics studies. Therefore this study aimed to determine the best methodology for the identification of the large diversity of lactobacilli species found in the vagina by comparing two techniques, one based on their biochemical profile and other employing molecular biology. A carbohydrate fermentation test (API 50 CH) was compared with multiplex polymerase chain reaction (PCR) for the identification of species of vaginal lactobacilli from 135 healthy women. The kappa index was used to evaluate agreement between the methods. Using the molecular technique, L. crispatus (32.6%), L. jensenii (25%) and L. gasseri (20.6%) were the most frequent species. However, using the biochemical technique, the most frequent species were: L. acidophilus (34.8%), L. crispatus (27.2%) and L. fermentum (13%). Although L. acidophilus was the most frequent specie found by biochemical tests, no strain of this microorganism was detected by PCR. Agreement between the methods was low for identification of all the most common species. Although rates of L. crispatus detected were similar using both methods (32.6% and 27.2%), agreement between them was relatively low (kappa = 0.52). CONCLUSIONS: Our results confirmed the limitation of the biochemical method and the applicability of a previously published molecular method (Multiplex PCR) for the identification of lactobacilli in the vaginal tract, focusing on further necessity of its improvement for also targeting L. vaginalis and L. iners.

[Study on virulence factors associated with biofilm formation and phylogenetic groupings in Escherichia coli strains isolated from patients with cystitis]. / Estudo dos fatores de virulência associados à formação de biofilme e agrupamento filogenético em Escherichia coli isoladas de pacientes com cistite.

Escherichia coli samples isolated from female patients with cystitis were characterized with regard to the presence of virulence factors associated with biofilm formation and phylogenetic groupings. Polymerase chain reaction results demonstrated that all the samples were positive for the gene fimH (type 1 fimbriae), 91 for fliC (flagellins), 50 for papC (P fimbriae), 44 for kpsMTII (capsules) and 36 for flu (antigen 43). The results from assays to quantify the biofilm formation demonstrated that 44 samples produced biofilm on polystyrene microplates and 56 samples produced weak or no biofilm. We also confirmed that Escherichia coli samples were present in phylogenetic groups B2 and D.

Estudo dos fatores de virulência associados à formação de biofilme e agrupamento filogenético em Escherichia coli isoladas de pacientes com cistite / Study on virulence factors associated with biofilm formation and phylogenetic groupings in Escherichia coli strains isolated from patients with cystitis

Amostras de Escherichia coli, isoladas de pacientes do sexo feminino com quadro clínico de cistite, foram caracterizadas quanto à presença de fatores de virulência associados à formação de biofilme e ao agrupamento filogenético. Os resultados da reação em cadeia da polimerase demonstraram que todas as amostras foram positivas para o gene fimH (fímbria do tipo1), 91 amostras foram positivas para o gene fliC (flagelina) 50 amostras positivas para o gene papC (fímbria P), 44 amostras positivas para o gene kpsMTII (cápsula) e 36 amostras positivas para o gene flu (antígeno 43). Os resultados dos ensaios de quantificação da formação de biofilme demonstraram que 44 amostras formaram biofilme em microplacas de poliestireno e 56 amostras apresentaram resultado ausente/fraco. Também confirmamos a incidência das amostras de Escherichia coli no grupo filogenético B2 e D.

Escherichia coli samples isolated from female patients with cystitis were characterized with regard to the presence of virulence factors associated with biofilm formation and phylogenetic groupings. Polymerase chain reaction results demonstrated that all the samples were positive for the gene fimH (type 1 fimbriae), 91 for fliC (flagellins), 50 for papC (P fimbriae), 44 for kpsMTII (capsules) and 36 for flu (antigen 43). The results from assays to quantify the biofilm formation demonstrated that 44 samples produced biofilm on polystyrene microplates and 56 samples produced weak or no biofilm. We also confirmed that Escherichia coli samples were present in phylogenetic groups B2 and D.

Genotypic characterization of virulence factors in Escherichia coli strains from patients with cystitis.

Adhesins (P-fimbriae, S-fimbriae, type 1 fimbriae and afimbrial adhesin), toxins (alpha-hemolysin and cytotoxic necrotizing factor type 1), iron acquisition systems (aerobactin) and host defense avoidance mechanisms (capsule or lipopolysaccharide) have been shown to be prevalent in Escherichia coli strains associated with urinary tract infections. In this work, 162 Uropathogenic Escherichia coli (UPEC) strains from patients with cystitis were genotypically characterized by polymerase chain reaction (PCR) assay. We developed three multiplex PCR assays for virulence-related genes papC, papE/F, papG alleles, fimH, sfa/foc, afaE, hly, cnf-1, usp, cdtB, iucD, and kpsMTII, all of them previously identified in UPEC strains. The PCR assay results identified 158 fimH (97.5%), 86 kpsMTII (53.1%), 53 papC/papEF/papG (32.7%), 45 sfa (27.8%), 42 iucD (25.9%), 41 hly (25.3%), 36 usp (22.2%), 30 cnf-1(18.5%) and 10 afa (6.2%) strains. No strain was positive for cdtB. In this work, we also demonstrated that adhesins may be multiple within a single strain and that several virulence genes can occur combined in association.

Genotypic characterization of virulence factors in Escherichia coli strains from patients with cystitis / Caracterização genotípica dos fatores de virulência em amostras de Escherichia coli isoladas de pacientes com cistite

Adhesins (P-fimbriae, S-fimbriae, type 1 fimbriae and afimbrial adhesin), toxins (α-hemolysin and cytotoxic necrotizing factor type 1), iron acquisition systems (aerobactin) and host defense avoidance mechanisms (capsule or lipopolysaccharide) have been shown to be prevalent in Escherichia coli strains associated with urinary tract infections. In this work, 162 Uropathogenic Escherichia coli (UPEC) strains from patients with cystitis were genotypically characterized by polymerase chain reaction (PCR) assay. We developed three multiplex PCR assays for virulence-related genes papC, papE/F, papG alleles, fimH, sfa/foc, afaE, hly, cnf-1, usp, cdtB, iucD, and kpsMTII, all of them previously identified in UPEC strains. The PCR assay results identified 158 fimH (97.5 percent), 86 kpsMTII (53.1 percent), 53 papC/papEF/papG (32.7 percent), 45 sfa (27.8 percent), 42 iucD (25.9 percent), 41 hly (25.3 percent), 36 usp (22.2 percent), 30 cnf-1(18.5 percent) and 10 afa (6.2 percent) strains. No strain was positive for cdtB. In this work, we also demonstrated that adhesins may be multiple within a single strain and that several virulence genes can occur combined in association.

Adesinas (Fímbria P, fímbria S, fímbria do tipo 1 e a adesina afimbrial), toxinas (α-hemolisina e o fator necrosante citotóxico do tipo 1), sistemas de captação de ferro (aerobactina), e mecanismos de defesa do hospedeiro (cápsula ou lipopolissacarídeo) são prevalentes em amostras de Escherichia coli associadas a infecções do trato urinário. O objetivo deste trabalho foi caracterizar genotipicamente 162 amostras de Escherichia coli uropatogênica (UPEC) de pacientes com cistite através do ensaio da reação em cadeia da polimerase. Foram realizados três ensaios de PCR multiplex para os seguintes fatores de virulência: papC, papE/F, alelos de papG, fimH, sfa/foc, afaE, hly, cnf-1, usp, cdtB, iucD, e kpsMTII. Os resultados da PCR identificaram, 158 amostras fimH (97,5 por cento), 86 amostras kpsMTII (53,1 por cento), 53 amostras papC/papEF/papG (32,7 por cento), 45 amostras sfa (27,8 por cento), 42 amostras iucD (25,9 por cento), 41 amostras hly (25,3 por cento), 36 amostras usp (22,2 por cento), 30 amostras cnf-1 (18,5 por cento) e 10 amostras afa (6,2 por cento). Nenhuma amostra foi positiva para o gene cdtB. Neste trabalho, demonstramos que podemos encontrar múltiplas adesinas em uma única amostra e que diferentes genes de fatores de virulência podem ser encontrados em associação.