Análise da virulência e resistência a antimicrobianos em linhagens de Staphylococcus spp. através de técnicas de genômica e proteômica
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2022-02-24
Autores
Silva, Nayara de Oliveira Gonçalves da
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Universidade Estadual Paulista (Unesp)
Resumo
O avanço das técnicas moleculares nas últimas décadas nos mostra que o genoma procarioto está em constante evolução. Tais mudanças podem resultar em alterações significativas no comportamento epidemiológico de diversas espécies de micro-organismos patogênicos. Staphylococcus aureus, principal representante do grupo dos coagulase-positivo, e Staphylococcus epidermidis, principal espécie no grupo dos estafilococos coagulase-negativos (ECN) persistem entre os principais patógenos causadores de infecções adquiridas em hospitais e na comunidade. Diante deste cenário, a virulência estafilocócica e a disseminação de linhagens multidroga resistentes (MDR) representam um sério problema de saúde pública. Neste contexto, alguns métodos são de extrema importância, como por exemplo a identificação e investigação molecular de genes e proteínas relacionados a patogenicidade. Além disso, o número crescente de sequências genômicas e proteômicas depositadas nos bancos de dados tem nos permitido um melhor entendimento acerca da variabilidade do perfil patogênico destes micro-organismos, permitindo assim elucidar os mecanismos relacionados a virulência, resistência e disseminação. Estudos envolvendo linhagens clínicas de ECN têm revelado a presença de genes relacionados a virulência e resistência em linhagens de S. epidermidis, evidenciando seu potencial patogênico, além da sua importância como um reservatório de genes, que podem ser transferidos para outras bactérias presentes em um mesmo nicho ecológico. O genoma e proteoma de S. aureus vêm sendo amplamente descritos, porém ainda mal caracterizados em linhagens de ECN. Tendo em vista a rápida evolução da virulência e resistência destes micro-organismos e seus altos índices de infecção e mortalidade, são necessários estudos que envolvam o sequenciamento do genoma e proteoma de linhagens de Staphylococcus spp., sobretudo de S. epidermidis, que vem se destacando como um importante patógeno emergente dentre os ECNs. Diante disso, investigamos linhagens de S. epidermidis isoladas de hemoculturas provenientes de recém-nascidos internados em uma Unidade de Terapia Intensiva Neonatal (UTIN). Foi realizado o sequenciamento do genoma e proteoma destas linhagens clínicas de S. epidermidis com o intuito de caracterizar a presença de genes e proteínas relacionados a patogenicidade destes isolados clínicos. Além disso, as amostras foram analisadas quanto ao perfil fenotípico de susceptibilidade a antimicrobianos. O resultado do sequenciamento do genoma total das duas linhagens clínicas revelou genes relacionados a resistência a tetraciclina, gentamicina, cefoxitina/oxacilina, penicilina, trimetoprim, ácido fusídico, ciprofloxacina e outras quinolonas, bem como genes codificadores de fatores de virulência para produção de enzimas e toxinas relacionados a mecanismos de aderência, evasão imune e sistemas de secreção. A análise proteômica mostrou a produção de proteínas relacionadas a patogênese, como por exemplo as enzimas SspA (serina V8 protease) e SspB (cisteína protease), além da autolisina Atl, validando os resultados obtidos a partir do sequenciamento. Os resultados deste estudo revelaram uma série de fatores de virulência que provavelmente contribuem para a patogenicidade de S. epidermidis, principalmente no que diz respeito a formação de biofilme, além de um repertório de genes de resistência aos antimicrobianos, alertando para a importância de S. epidermidis como patógenos emergentes no contexto das infecções nosocomiais. Tendo em vista que as populações de S. aureus e S. epidermidis podem ocupar nichos ecológicos sobrepostos, o fluxo gênico pode ocorrer entre essas espécies, o que destaca a importância da vigilância contínua para detectar novas linhagens recombinantes que promovem a emergência de micro-organismos multirresistentes e virulentos.
The advancement of molecular techniques in recent decades shows us that the prokaryotic genome is constantly evolving. Such changes can result in significant alterations in the epidemiological behavior of several species of pathogenic microorganisms. main representative of the coagulasepositive group, and Staph ylococcus aureus Staphylococcus epidermidis , the , the main species of the coagulasenegative staphylococci ( that cause infections acquired in hospitals and S CN) group persist among the main pathogens in the community. In this scenario, staphylococcal virulence and the spread of multidrug resistant (MDR) strains represent a serious public health problem. In this context, some methods are extremely important, such as the identification and molecular inv estigation of genes and proteins related to pathogenicity. In addition, the increasing number of genomic and proteomic sequences deposited in databases has allowed us to better understand the variability of the pathogenic profile of these microorganisms, t hus allowing us to elucidate the mechanisms related to virulence, resistance and dissemination. Studies involving clinical strains of and resistance in strains of SCN have revealed the presence of genes related to virulence S. epidermidis , evidencing its p athogenic potential, in addition to its importance as a reservoir of genes, which can be transferred to other bacteria. present in the same ecological niche. The genome and proteome of still poorly characterized in SCN S. aureus have been widely described, but strains. In view of the rapid evolution of virulence and resistance of these microorganisms and their high rates of infection and mortality, studies involving the sequencing of the genome and proteome of strains of epidermidis Staphylococcus spp., especially , which has been standing out as an important emerging pathogen among S. SCN s. Therefore, we investigated S. epidermidis strains isolated from blood cultures from newborns admitted to a N eonatal these clinical strains of I ntensive C are S. epidermidis U nit (NICU). The genome and proteome s equencing of was performed in order to characterize the presence of genes and proteins related to the pathogenicity of these clinical isolates. Furthermore, the samples were analyzed for the phenotypic profile of an timicrobial susceptibility. The result of whole genome sequencing of the two clinical strains revealed genes related to resistance to tetracycline, gentamicin, cefoxitin/oxacillin, penicillin, trimethoprim, fusidic acid, ciprofloxacin and other quinolones, as well as genes encoding virulence factors for production of enzymes and toxins related to adherence mechanisms, immune evasion and secretion systems. The proteomic analysis showed the production of proteins related to pathogenesis, such as the enzymes S spA (serine V8 protease) and SspB (cysteine protease), in addition to autolysin Atl, validating the results obtained from the sequencing. The results of this study revealed a series of virulence factors that probably contribute to the pathogenicity of to biofilm formation, in addition to a S. epidermidis , especially with regard repertoire of antimicrobial resistance genes, alerting to the importance of S. epidermidis Considering that the popul as emerging pathogens in the context of nosocomial infections. ations of S. aureus and S. epidermidis can occupy overlapping ecological niches, gene flow can occur between these species, which highlights the importance of continuous surveillance to detect new recombinant lineages that promote the emergence of multidru gresistant and virulent microorganisms.
The advancement of molecular techniques in recent decades shows us that the prokaryotic genome is constantly evolving. Such changes can result in significant alterations in the epidemiological behavior of several species of pathogenic microorganisms. main representative of the coagulasepositive group, and Staph ylococcus aureus Staphylococcus epidermidis , the , the main species of the coagulasenegative staphylococci ( that cause infections acquired in hospitals and S CN) group persist among the main pathogens in the community. In this scenario, staphylococcal virulence and the spread of multidrug resistant (MDR) strains represent a serious public health problem. In this context, some methods are extremely important, such as the identification and molecular inv estigation of genes and proteins related to pathogenicity. In addition, the increasing number of genomic and proteomic sequences deposited in databases has allowed us to better understand the variability of the pathogenic profile of these microorganisms, t hus allowing us to elucidate the mechanisms related to virulence, resistance and dissemination. Studies involving clinical strains of and resistance in strains of SCN have revealed the presence of genes related to virulence S. epidermidis , evidencing its p athogenic potential, in addition to its importance as a reservoir of genes, which can be transferred to other bacteria. present in the same ecological niche. The genome and proteome of still poorly characterized in SCN S. aureus have been widely described, but strains. In view of the rapid evolution of virulence and resistance of these microorganisms and their high rates of infection and mortality, studies involving the sequencing of the genome and proteome of strains of epidermidis Staphylococcus spp., especially , which has been standing out as an important emerging pathogen among S. SCN s. Therefore, we investigated S. epidermidis strains isolated from blood cultures from newborns admitted to a N eonatal these clinical strains of I ntensive C are S. epidermidis U nit (NICU). The genome and proteome s equencing of was performed in order to characterize the presence of genes and proteins related to the pathogenicity of these clinical isolates. Furthermore, the samples were analyzed for the phenotypic profile of an timicrobial susceptibility. The result of whole genome sequencing of the two clinical strains revealed genes related to resistance to tetracycline, gentamicin, cefoxitin/oxacillin, penicillin, trimethoprim, fusidic acid, ciprofloxacin and other quinolones, as well as genes encoding virulence factors for production of enzymes and toxins related to adherence mechanisms, immune evasion and secretion systems. The proteomic analysis showed the production of proteins related to pathogenesis, such as the enzymes S spA (serine V8 protease) and SspB (cysteine protease), in addition to autolysin Atl, validating the results obtained from the sequencing. The results of this study revealed a series of virulence factors that probably contribute to the pathogenicity of to biofilm formation, in addition to a S. epidermidis , especially with regard repertoire of antimicrobial resistance genes, alerting to the importance of S. epidermidis Considering that the popul as emerging pathogens in the context of nosocomial infections. ations of S. aureus and S. epidermidis can occupy overlapping ecological niches, gene flow can occur between these species, which highlights the importance of continuous surveillance to detect new recombinant lineages that promote the emergence of multidru gresistant and virulent microorganisms.
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Palavras-chave
Staphylococcus epidermidis, Virulência, Unidade de Terapia Intensiva Neonatal, Resistência a antimicrobianos, Antimicrobial resistance, Virulence, Neonatal Intensive Care Unit.