Determinação da relação clonal e virulência de Staphylococcus aureus isolados de pacientes vivendo com HIV/AIDS e seus familiares
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Data
2018-05-18
Autores
Souza, Camila Sena Martins de [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Indivíduos colonizados podem contribuir para disseminação de Staphylococcus aureus, que se destaca por sua patogenicidade e alta frequência, capaz de produzir doenças tanto em indivíduos sadios quanto em imunocomprometidos, sendo esse risco aumentado na imunodeficiência humana (HIV/aids). Esse estudo teve como objetivos determinar a relação clonal de MRSA (Staphylococcus aureus Resistente à Meticilina) e MSSA (Staphylococcus aureus Sensível à Meticilina) entre as cepas isoladas de pessoas vivendo com HIV/aids (PVHA) e seus contactantes domiciliares, e os fatores de virulência que podem contribuir para o carreamento desses micro-organismos. S. aureus foram isolados da nasofaringe e orofaringe de 368 PVHA do Serviço de Ambulatórios Especializados de Infectologia “Domingos Alves Meira” (SAE/DAM) do complexo da Faculdade de Medicina de Botucatu (FMB) e Fundação para o Desenvolvimento Médico Hospitalar (FAMESP), sendo 112 residentes em Botucatu e 256 provenientes de cidades da região. A técnica de PCR (Reação em Cadeia da Polimerase) foi utilizada para detecção do gene mecA, genes das enterotoxinas (sea, seb, e sec-1), esfoliatinas A e B (eta e etb), toxina 1 da síndrome do choque tóxico (tst), leucocidina Panton–Valentine (lukS-PV e lukF-PV), hemolisinas alfa e delta (hla and hld) e biofilme (icaA e icaD) e para tipagem do SCCmec. Para a tipagem molecular foi utilizado a técnica de Pulsed Field Gel Electrophoresis (PFGE), Multilocus Sequence Typing (MLST) e spa Typing. Os pacientes residentes em Botucatu foram convidados a mais duas coletas com inclusão dos contactantes domiciliares, totalizando 73 contactantes. Além disso, foi realizado o georreferenciamento dos domicílios no momento da coleta da área urbana de Botucatu. A prevalência de colonização por S. aureus na primeira coleta foi de 26%, com dez indivíduos colonizados com MRSA, com a taxa de prevalência de 2,7%. Na segunda coleta a prevalência foi de 23,8%, com a presença de três isolados de MRSA, já entre os contactantes domiciliares foi de 30%, com a identificação de quatro MRSA. E por fim, terceira e última coleta a prevalência encontrada foi de 25,3% com dois MRSA. Do total de 178 isolados de S. aureus, 19 (10,6%) eram carreadores do gene mecA, dentre esses, quatro eram provenientes de contactantes domiciliares, com prevalência do SCCmec Tipo IV. A identificação do perfil clonal através da análise por PFGE evidenciou a presença de 2 clusters majoritários entre os MRSA e 10 clusters entre os isolados sensíveis, demonstrando possíveis transmissões no ambiente familiar. Quando realizada análise pelo MSLT identificou-se a presença de um mesmo clone em cidades distintas sugerindo ampla disseminação do clone no Estado de São Paulo. Encontramos S. aureus isolados dos pacientes bem distribuídos em toda a cidade de Botucatu, no entanto, os isolados MRSA não foram encontrados na região central da cidade. Além disso, os resultados demonstraram uma prevalência significante de genes de virulência nos isolados, com destaque para genes das hemolisinas e genes para produção de biofilme. Em relação ao georreferenciamento, observamos uma distribuição de S. aureus em toda a cidade de Botucatu, porém, a prevalência maior de MRSA encontra-se na região norte da cidade. A análise do perfil clonal leva a maiores discussões sobre o processo de evolução da bactéria, sugerindo uma provável origem comum entre as cepas de diferentes cidades.
Colonized individuals may contribute to the dissemination of Staphylococcus aureus, which stands out for its pathogenicity and high frequency, capable of producing diseases in both healthy and immunocompromised individuals, with an increased risk of human immunodeficiency (HIV/AIDS). This study aimed to determine the clonal relationship between MRSA (Methicillin Resistant Staphylococcus aureus) and MSSA (Methicillin-Sensitive Staphylococcus aureus) between strains isolated from people living with HIV/aids (PLHA) and their home contacts, and virulence factors which may contribute to the transport of these microorganisms. S. aureus were isolated from the nasopharynx and oropharynx of 368 PLHA from the Specialized Outpatient Services “Domingos Alves Meira” (SAE/DAM) of the Botucatu Medical School (FMB) and Hospital Medical Development Foundation (FAMESP), of which 112 were residents in Botucatu and 256 from cities in the region. The PCR (Polymerase Chain Reaction) technique was used to detect the mecA gene, enterotoxin genes (sea, seb, and sec-1), esfoliatins A and B (eta and etb), toxic shock syndrome toxin 1 (tst), Panton-Valentine leukocidin (lukS-PV and lukF-PV), alpha and delta hemolysins (hla and hld) and biofilm (icaA and icaD) and for typing SCCmec. For molecular typing were used techniques of the Pulsed Field Gel Electrophoresis (PFGE), Multilocus Sequence Typing (MLST) and spa Typing. The patients residing in Botucatu were invited to more two collections with the inclusion of the home contacts, totaling 73 home contacts. In addition, the georeferencing of households was carried out at the time of collection of the urban area of Botucatu. The prevalence of S. aureus colonization in the first collection was 26%, with ten individuals colonized with MRSA, with a prevalence rate of 2.7%. In the second collection the prevalence was 23.8%, with the presence of three MRSA isolates; among the home contacts, 30% were identified, with the identification of four MRSAs. Finally, the third and final collection was found to be 25.3% with two MRSA. Of the total of 178 S. aureus isolates, 19 (10.6%) were carriers of the mecA gene, of which four were from home contacts, with prevalence of SCCmec Type IV. The identification of the clonal profile through PFGE analysis evidenced the presence of two major clusters between MRSA and 10 clusters among the sensitive isolates, demonstrating possible transmissions in the family environment. When MSLT analysis was performed, the presence of a same clone was identified in distinct cities, suggesting a wide dissemination of the clone in the State of São Paulo. We found S. aureus isolated from patients well distributed throughout the city of Botucatu, however, MRSA isolates were not found in the central region of the city. In addition, the results demonstrated a significant prevalence of virulence genes in the isolates, with emphasis on hemolysin genes and genes for biofilm production. Regarding georeferencing, we observed a distribution of S. aureus throughout the city of Botucatu, however, the highest prevalence of MRSA is found in the northern region of the city. The analysis of the clonal profile leads to further discussions about the process of evolution of the bacteria, suggesting a probable common origin among the strains of different cities.
Colonized individuals may contribute to the dissemination of Staphylococcus aureus, which stands out for its pathogenicity and high frequency, capable of producing diseases in both healthy and immunocompromised individuals, with an increased risk of human immunodeficiency (HIV/AIDS). This study aimed to determine the clonal relationship between MRSA (Methicillin Resistant Staphylococcus aureus) and MSSA (Methicillin-Sensitive Staphylococcus aureus) between strains isolated from people living with HIV/aids (PLHA) and their home contacts, and virulence factors which may contribute to the transport of these microorganisms. S. aureus were isolated from the nasopharynx and oropharynx of 368 PLHA from the Specialized Outpatient Services “Domingos Alves Meira” (SAE/DAM) of the Botucatu Medical School (FMB) and Hospital Medical Development Foundation (FAMESP), of which 112 were residents in Botucatu and 256 from cities in the region. The PCR (Polymerase Chain Reaction) technique was used to detect the mecA gene, enterotoxin genes (sea, seb, and sec-1), esfoliatins A and B (eta and etb), toxic shock syndrome toxin 1 (tst), Panton-Valentine leukocidin (lukS-PV and lukF-PV), alpha and delta hemolysins (hla and hld) and biofilm (icaA and icaD) and for typing SCCmec. For molecular typing were used techniques of the Pulsed Field Gel Electrophoresis (PFGE), Multilocus Sequence Typing (MLST) and spa Typing. The patients residing in Botucatu were invited to more two collections with the inclusion of the home contacts, totaling 73 home contacts. In addition, the georeferencing of households was carried out at the time of collection of the urban area of Botucatu. The prevalence of S. aureus colonization in the first collection was 26%, with ten individuals colonized with MRSA, with a prevalence rate of 2.7%. In the second collection the prevalence was 23.8%, with the presence of three MRSA isolates; among the home contacts, 30% were identified, with the identification of four MRSAs. Finally, the third and final collection was found to be 25.3% with two MRSA. Of the total of 178 S. aureus isolates, 19 (10.6%) were carriers of the mecA gene, of which four were from home contacts, with prevalence of SCCmec Type IV. The identification of the clonal profile through PFGE analysis evidenced the presence of two major clusters between MRSA and 10 clusters among the sensitive isolates, demonstrating possible transmissions in the family environment. When MSLT analysis was performed, the presence of a same clone was identified in distinct cities, suggesting a wide dissemination of the clone in the State of São Paulo. We found S. aureus isolated from patients well distributed throughout the city of Botucatu, however, MRSA isolates were not found in the central region of the city. In addition, the results demonstrated a significant prevalence of virulence genes in the isolates, with emphasis on hemolysin genes and genes for biofilm production. Regarding georeferencing, we observed a distribution of S. aureus throughout the city of Botucatu, however, the highest prevalence of MRSA is found in the northern region of the city. The analysis of the clonal profile leads to further discussions about the process of evolution of the bacteria, suggesting a probable common origin among the strains of different cities.
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Palavras-chave
Staphylococcus aureus, fatores de virulência, pessoa vivendo com HIV/aids, PFGE, MLST