Similaridade genética pelo RAPD-PCR de cepas de Staphylococcus sp isolados de portadores humanos e de camas de uma unidade hospitalar
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Data
2007-12-10
Autores
Fabiano, Telma Luciana Trovó [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Foram coletadas 143 amostras de mãos de humanos e camas hospitalares, através de suabes em caldo BHI (Brain Heart Infusion), em um Hospital Escola de Ribeirão Preto, SP. Este trabalho teve como objetivo caracterizar as cepas de Staphylococcus sp e verificar o grau de similaridade genética entre as cepas pelo RAPD-PCR e analisar o perfil de resistência a diversos antimicrobianos. As amostras coletadas foram incubadas a 37º C por 24 horas e após este período as culturas foram semeadas em placas de Petri contendo agar “Staphylococcus Médium 110”. As colônias típicas do gênero Staphylococcus foram coletadas e estocadas a 4º C até o momento de elaboração das provas de produção de catalase e coagulase, fermentação do manitol, DNAse e hemólise. As cepas isoladas foram analisadas através da técnica de RAPD-PCR para verificar o grau de similaridade. A sensibilidade das cepas isoladas foi testada frente a 11 diferentes antibióticos. Foram isoladas 92 cepas de Staphylococcus sp sendo 67 (72,8%) cepas de Staphylococcus coagulase negativas e 25 (27,2%) cepas de Staphylococcus coagulase positivas. A análise de similaridade mostrou heterogeneidade genética entre as cepas. Foram encontradas oito (8.7%) cepas de Staphylococcus sp resistentes à vancomicina, sendo uma multirresistente a todos os antibióticos. Introduzir medidas de assepsia nas mãos de pessoal e leitos hospitalares e a racionalização do uso indiscriminado de antibióticos muito contribuirá para diminuição das infecções nosocomiais.
A total of 143 samples were analyzed in this study. These included samples from human hands and hospital beds at a Medical Teaching Hospital of Ribeirão Preto, SP. This study aimed to characterize the strains of Staphylococcus sp and verify the degree of genetic similarity between the strains by RAPD-PCR and analyse the profile of the various antimicrobial resistance. Swabs samples were collected and cultured in brain heart infusion medium at 37o C for 24 h. Further cultivation was performed on Staphylococcus medium 110 agar plates. Typical colonies representing Staphylococcus genus were collected and stored at 4º C until it was used for identification tests, which included catalase, coagulase and mannitol production, hemolysis and Dnase. Isolates were analyzed by RAPD-PCR to verify similarity level. Antibiotic susceptibility was determined using 11 different antibiotics. A total of 92 Staphylococcus sp strains were isolated, whereas 67 (72.8%) were coagulase-negative Staphylococcus strains and 25 (27,2%) were coagulase-positive Staphylococcus strains. Similarity analysis revealed a great dissimilarity among isolates. High percentage of beta-lactams resistant Staphylococcus sp was found. Eight strains (8.7%) were vancomycin-resistant. A high percentage of multiresistant strains were found. Introduce of good hygiene practices for hands and hospital beds and adequate antibiotic use, may contribute to reduction of nosocomial infection and consequently improved antibiotic treatment response.
A total of 143 samples were analyzed in this study. These included samples from human hands and hospital beds at a Medical Teaching Hospital of Ribeirão Preto, SP. This study aimed to characterize the strains of Staphylococcus sp and verify the degree of genetic similarity between the strains by RAPD-PCR and analyse the profile of the various antimicrobial resistance. Swabs samples were collected and cultured in brain heart infusion medium at 37o C for 24 h. Further cultivation was performed on Staphylococcus medium 110 agar plates. Typical colonies representing Staphylococcus genus were collected and stored at 4º C until it was used for identification tests, which included catalase, coagulase and mannitol production, hemolysis and Dnase. Isolates were analyzed by RAPD-PCR to verify similarity level. Antibiotic susceptibility was determined using 11 different antibiotics. A total of 92 Staphylococcus sp strains were isolated, whereas 67 (72.8%) were coagulase-negative Staphylococcus strains and 25 (27,2%) were coagulase-positive Staphylococcus strains. Similarity analysis revealed a great dissimilarity among isolates. High percentage of beta-lactams resistant Staphylococcus sp was found. Eight strains (8.7%) were vancomycin-resistant. A high percentage of multiresistant strains were found. Introduce of good hygiene practices for hands and hospital beds and adequate antibiotic use, may contribute to reduction of nosocomial infection and consequently improved antibiotic treatment response.
Descrição
Palavras-chave
Estafilococos, Índice de similaridade, RAPD-PCR, Multirresistência, Similary index, Multiresistance, Staphylococcus
Como citar
FABIANO, Telma Luciana Trovó. Similaridade genética pelo RAPD-PCR de cepas de Staphylococcus sp isolados de portadores humanos e de camas de uma unidade hospitalar. 2007. xii, 63 f. Tese (doutorado) - Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, 2007.