Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase

Detalhes bibliográficos
Ano de defesa: 2018
Autor(a) principal: Maristela Oliveira Lara
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Minas Gerais
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: https://hdl.handle.net/1843/BUOS-B46H9Z
Resumo: Healthcare-associated infections (HAIs) are worrying adverse events in public health. They are an important cause of morbidity and mortality in intensive care units. Invasive devices such as the central venous catheter (CVC) favors a type of HAIs, the bloodstream infection. This event is commonly diagnosed by blood culture and/or culture of the catheter tip, however, the response time of these tests or their results not always contribute to the appropriate treatment. Advances in biotechnology provide tools capable of contributing to diagnoses of infection. The aim of the present thesis was to detect potentially pathogenic bacteria at the tip of a central venous catheter through polymerase chain reaction (PCR). Subjects were treated with DNA extraction and molecular tracing in CVC. It is a cross-sectional molecular study. Laboratory tests comparing DNA extraction methods were performed with the Staphylococcus aureus bacterium for subsequent application to catheters collected from patients. Over a period of 6 months, in an Adult Intensive Care Unit of a philanthropic and training hospital, (n=34) catheters were removed from patients under suspicion of bloodstream infection. All the thirty-four catheters were subjected to DNA extraction from the biological material contained in their wall and inside their lumens. The bacteria were identified by PCR using a standard set of reagents and temperatures. The results found in the analysis by molecular biology were compared with the results of the cultures of these patients, performed by the hospital. Collection of patients' data was also carried out: sex, age, use of other invasive devices, CVC insertion location and period of catheters use; and presence of phlogistic signs in the insertion site of the device. Statistical tests were used with the help of the Stata software, version 15. The prevalence of bacteria in CVCs was: Staphylococcus aureaus (50%), Enterococcus faecalis (41,2%), Klebsiella pneumoniae (32,4%), Pseudomonas aeruginosa (20,6%), Acinetobacter baumannii (38,2%) and Escherichia coli (2,9%). All blood cultures performed had no bacteria as a result of the examination. Catheter-tip culture revealed microorganisms in 21 (61.8%) devices, whereas PCR showed positivity in 31 (91.2%). The most commonly detected pathogens are usually found in the environment and in the microbioma of the skin and they are possibly transmitted to patients by the hands of health professionals. These findings are relevant when programming CVC-related bloodstream infection prevention measures. The genomic material extraction method, primers panel and amplification protocol of this study identified the major pathogens prevalent in bloodstream infections. In this way, molecular identification of bacteria may assist in the detection of bloodstream infection and decision-making regarding the choice of the best therapy.
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spelling 2019-08-12T07:30:28Z2025-09-08T23:25:46Z2019-08-12T07:30:28Z2018-04-03https://hdl.handle.net/1843/BUOS-B46H9ZHealthcare-associated infections (HAIs) are worrying adverse events in public health. They are an important cause of morbidity and mortality in intensive care units. Invasive devices such as the central venous catheter (CVC) favors a type of HAIs, the bloodstream infection. This event is commonly diagnosed by blood culture and/or culture of the catheter tip, however, the response time of these tests or their results not always contribute to the appropriate treatment. Advances in biotechnology provide tools capable of contributing to diagnoses of infection. The aim of the present thesis was to detect potentially pathogenic bacteria at the tip of a central venous catheter through polymerase chain reaction (PCR). Subjects were treated with DNA extraction and molecular tracing in CVC. It is a cross-sectional molecular study. Laboratory tests comparing DNA extraction methods were performed with the Staphylococcus aureus bacterium for subsequent application to catheters collected from patients. Over a period of 6 months, in an Adult Intensive Care Unit of a philanthropic and training hospital, (n=34) catheters were removed from patients under suspicion of bloodstream infection. All the thirty-four catheters were subjected to DNA extraction from the biological material contained in their wall and inside their lumens. The bacteria were identified by PCR using a standard set of reagents and temperatures. The results found in the analysis by molecular biology were compared with the results of the cultures of these patients, performed by the hospital. Collection of patients' data was also carried out: sex, age, use of other invasive devices, CVC insertion location and period of catheters use; and presence of phlogistic signs in the insertion site of the device. Statistical tests were used with the help of the Stata software, version 15. The prevalence of bacteria in CVCs was: Staphylococcus aureaus (50%), Enterococcus faecalis (41,2%), Klebsiella pneumoniae (32,4%), Pseudomonas aeruginosa (20,6%), Acinetobacter baumannii (38,2%) and Escherichia coli (2,9%). All blood cultures performed had no bacteria as a result of the examination. Catheter-tip culture revealed microorganisms in 21 (61.8%) devices, whereas PCR showed positivity in 31 (91.2%). The most commonly detected pathogens are usually found in the environment and in the microbioma of the skin and they are possibly transmitted to patients by the hands of health professionals. These findings are relevant when programming CVC-related bloodstream infection prevention measures. The genomic material extraction method, primers panel and amplification protocol of this study identified the major pathogens prevalent in bloodstream infections. In this way, molecular identification of bacteria may assist in the detection of bloodstream infection and decision-making regarding the choice of the best therapy.Universidade Federal de Minas GeraisCateterismo Venoso CentralInfecções Relacionadas a CateterDNA BacterianoReação em Cadeia da PolimeraseCateteresInfecção HospitalarInfecção hospitalarCateterismo venoso centralDNA bacterianoMedicinaInfecções relacionadas a cateterReação em cadeia da polimeraseDetecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimeraseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisMaristela Oliveira Larainfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGCarla Jorge MachadoMarcelo CarneiroJuliana Ladeira GarbaccioAdriana Cristina de OliveiraAs infecções relacionadas à assistência à saúde (IRAS) são eventos adversos preocupantes em saúde pública, que se configuram como importante causa de morbidade e mortalidade em unidades de terapia intensiva. Dispositivos invasivos como o cateter venoso central (CVC) favorecem um tipo de IRAS, a infecção da corrente sanguínea. Esse evento é comumente diagnosticado por hemocultura e ou cultura da ponta do cateter, entretanto nem sempre o tempo de resposta dos exames ou os achados contribuem com o adequado tratamento. Os avanços em biotecnologia apontam ferramentas capazes de contribuir com diagnósticos de infecção. O objetivo da presente tese foi testar a técnica de reação em cadeia da polimerase (PCR) como ferramenta para detecção de bactérias potencialmente patogênicas em ponta de CVC de pacientes com suspeita de infecção da corrente sanguínea relacionada ao cateter, internados na Unidade de Terapia Intensiva de Adultos de um hospital filantrópico e de ensino no interior de Minas Gerais. Foram abordados os temas extração de DNA e rastreamento molecular em CVC. Tratou-se de um estudo molecular, transversal, descritivo e exploratório. Testes laboratoriais de comparação entre métodos de extração de DNA foram realizados com cepa da bactéria Staphylococcus aureus para posterior aplicação em cateteres coletados de pacientes. Durante um período de seis meses, uma amostra de conveniência com trinta e quarto cateteres removidos de pacientes internados na Unidade de Terapia Intensiva de Adultos, sob suspeita de infecção da corrente sanguínea, foram submetidos à extração de DNA do material biológico contido na parede externa e no interior dos lúmens dos mesmos. Procedeu-se a identificação de bactérias por PCR utilizando um padrão de reagentes e temperaturas. Os resultados encontrados na análise por biologia molecular foram comparados com os resultados das culturas desses pacientes, realizadas pelo hospital. Houve ainda, o levantamento em prontuário de dados dos pacientes: sexo, idade, uso de outros dispositivos invasivos, tempo de permanência do CVC e local de inserção do cateter; e presença de sinais flogísticos no local de inserção do dispositivo. Testes estatísticos com auxílio do programa Stata, versão 15, foram utilizados. A prevalência das bactérias no CVC por teste de PCR foi: Staphylococcus aureaus (50%), Enterococcus faecalis (41,2%), Klebsiella pneumoniae (32,4%), Pseudomonas aeruginosa (20,6%), Acinetobacter baumannii (38,2%) e Escherichia coli (2,9%). Todas as hemoculturas realizadas tiveram ausência de bactérias como resultado do exame. A cultura de ponta de cateter revelou bactérias em 21 (61,8%) dispositivos, enquanto a PCR apresentou positividade em 31 (91,2%). Os patógenos mais detectados são comumente encontrados no ambiente e no microbioma humano, transmitidos aos pacientes inclusive pelas mãos dos profissionais de saúde. Estes achados são relevantes ao se programar medidas de prevenção de infecção da corrente sanguínea relacionada ao CVC. O método de extração do material genômico, o painel de primers e protocolo de amplificação deste estudo identificaram os principais bactérias comumente prevalentes nas infecções da corrente sanguínea. Desta forma, a identificação molecular de bactérias poderá auxiliar na detecção de infecção da corrente sanguínea e a tomada de decisão relativa à escolha da melhor terapia.UFMGORIGINALcapa_dura.pdfapplication/pdf4055153https://repositorio.ufmg.br//bitstreams/2b5974fa-587f-4d1f-b148-912322b4f313/download3310886c4e1c9ba0281f38f2bf9b98f2MD51trueAnonymousREADTEXTcapa_dura.pdf.txttext/plain124723https://repositorio.ufmg.br//bitstreams/759564fb-73a9-4170-99b1-59122c3ecbb7/downloadc9b7d89cc970998699d7aa89e4b46a78MD52falseAnonymousREAD1843/BUOS-B46H9Z2025-09-08 20:25:46.013open.accessoai:repositorio.ufmg.br:1843/BUOS-B46H9Zhttps://repositorio.ufmg.br/Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2025-09-08T23:25:46Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
title Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
spellingShingle Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
Maristela Oliveira Lara
Infecção hospitalar
Cateterismo venoso central
DNA bacteriano
Medicina
Infecções relacionadas a cateter
Reação em cadeia da polimerase
Cateterismo Venoso Central
Infecções Relacionadas a Cateter
DNA Bacteriano
Reação em Cadeia da Polimerase
Cateteres
Infecção Hospitalar
title_short Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
title_full Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
title_fullStr Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
title_full_unstemmed Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
title_sort Detecção de patógenos em ponta de cateter venoso central por reação em cadeia da polimerase
author Maristela Oliveira Lara
author_facet Maristela Oliveira Lara
author_role author
dc.contributor.author.fl_str_mv Maristela Oliveira Lara
dc.subject.por.fl_str_mv Infecção hospitalar
Cateterismo venoso central
DNA bacteriano
Medicina
Infecções relacionadas a cateter
Reação em cadeia da polimerase
topic Infecção hospitalar
Cateterismo venoso central
DNA bacteriano
Medicina
Infecções relacionadas a cateter
Reação em cadeia da polimerase
Cateterismo Venoso Central
Infecções Relacionadas a Cateter
DNA Bacteriano
Reação em Cadeia da Polimerase
Cateteres
Infecção Hospitalar
dc.subject.other.none.fl_str_mv Cateterismo Venoso Central
Infecções Relacionadas a Cateter
DNA Bacteriano
Reação em Cadeia da Polimerase
Cateteres
Infecção Hospitalar
description Healthcare-associated infections (HAIs) are worrying adverse events in public health. They are an important cause of morbidity and mortality in intensive care units. Invasive devices such as the central venous catheter (CVC) favors a type of HAIs, the bloodstream infection. This event is commonly diagnosed by blood culture and/or culture of the catheter tip, however, the response time of these tests or their results not always contribute to the appropriate treatment. Advances in biotechnology provide tools capable of contributing to diagnoses of infection. The aim of the present thesis was to detect potentially pathogenic bacteria at the tip of a central venous catheter through polymerase chain reaction (PCR). Subjects were treated with DNA extraction and molecular tracing in CVC. It is a cross-sectional molecular study. Laboratory tests comparing DNA extraction methods were performed with the Staphylococcus aureus bacterium for subsequent application to catheters collected from patients. Over a period of 6 months, in an Adult Intensive Care Unit of a philanthropic and training hospital, (n=34) catheters were removed from patients under suspicion of bloodstream infection. All the thirty-four catheters were subjected to DNA extraction from the biological material contained in their wall and inside their lumens. The bacteria were identified by PCR using a standard set of reagents and temperatures. The results found in the analysis by molecular biology were compared with the results of the cultures of these patients, performed by the hospital. Collection of patients' data was also carried out: sex, age, use of other invasive devices, CVC insertion location and period of catheters use; and presence of phlogistic signs in the insertion site of the device. Statistical tests were used with the help of the Stata software, version 15. The prevalence of bacteria in CVCs was: Staphylococcus aureaus (50%), Enterococcus faecalis (41,2%), Klebsiella pneumoniae (32,4%), Pseudomonas aeruginosa (20,6%), Acinetobacter baumannii (38,2%) and Escherichia coli (2,9%). All blood cultures performed had no bacteria as a result of the examination. Catheter-tip culture revealed microorganisms in 21 (61.8%) devices, whereas PCR showed positivity in 31 (91.2%). The most commonly detected pathogens are usually found in the environment and in the microbioma of the skin and they are possibly transmitted to patients by the hands of health professionals. These findings are relevant when programming CVC-related bloodstream infection prevention measures. The genomic material extraction method, primers panel and amplification protocol of this study identified the major pathogens prevalent in bloodstream infections. In this way, molecular identification of bacteria may assist in the detection of bloodstream infection and decision-making regarding the choice of the best therapy.
publishDate 2018
dc.date.issued.fl_str_mv 2018-04-03
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