Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos

Detalhes bibliográficos
Autor(a) principal: Fernandes, Tássia Péres
Data de Publicação: 2022
Tipo de documento: Trabalho de conclusão de curso
Idioma: por
Título da fonte: Repositório Institucional da UFAM
Texto Completo: http://riu.ufam.edu.br/handle/prefix/6237
Resumo: In the last years, the reports of polymyxins resistance have increased considerably, so efficient, reliable, fast and easily interpreted testing to determine polymyxins susceptibility is necessary. The present study aimed to develop and evaluate the performance of a colorimetric microelution method for detecting polymyxin B resistance in clinical isolates of Gram-negative bacilli. The tests were performed with 11 clinical isolates, including 8 E. coli and 3 K. pneumoniae. The colorimetric polymyxin B microelution method was an adaptation of the colistin broth disk elution test, adding resazurin at the end of the process. The results were interpreted using the breakpoints of the BrCAST. When evaluating the microelution results with and without resazurin, 7 isolates presented categorical agreement and 9 and 10 isolates, respectively, presented essential agreement with the reference method. In conclusion, both polymyxin B microelution test and the colorimetric polymyxin B microelution test were easy to perform, requiring materials commonly found in routine microbiology laboratories. Furthermore, a method involving polymyxin B, which is the polymyxin used in Brazil, encourages the study of new susceptibility tests against polymyxins.
id UFAM-1_fde9a594f930585d0958df4098fcceae
oai_identifier_str oai:localhost:prefix/6237
network_acronym_str UFAM-1
network_name_str Repositório Institucional da UFAM
repository_id_str
spelling Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativosEnterobacteriaceaePolimixina B.PolimixinasResistência bacteriana a antibióticosCIÊNCIAS DA SAÚDEResazurinaCalorimetria de microeluiçãoPropriedade policatiônicaMicroeluição da colistina em placaAcinetobacter spp.In the last years, the reports of polymyxins resistance have increased considerably, so efficient, reliable, fast and easily interpreted testing to determine polymyxins susceptibility is necessary. The present study aimed to develop and evaluate the performance of a colorimetric microelution method for detecting polymyxin B resistance in clinical isolates of Gram-negative bacilli. The tests were performed with 11 clinical isolates, including 8 E. coli and 3 K. pneumoniae. The colorimetric polymyxin B microelution method was an adaptation of the colistin broth disk elution test, adding resazurin at the end of the process. The results were interpreted using the breakpoints of the BrCAST. When evaluating the microelution results with and without resazurin, 7 isolates presented categorical agreement and 9 and 10 isolates, respectively, presented essential agreement with the reference method. In conclusion, both polymyxin B microelution test and the colorimetric polymyxin B microelution test were easy to perform, requiring materials commonly found in routine microbiology laboratories. Furthermore, a method involving polymyxin B, which is the polymyxin used in Brazil, encourages the study of new susceptibility tests against polymyxins.Nos últimos anos a resistência as polimixinas têm aumentado consideravelmente, portanto é de extrema importância métodos eficientes, confiáveis, rápidos e de fácil interpretação para determinar a suscetibilidade às polimixinas. O presente estuda objetivou desenvolver e avaliar o desempenho de um método de microeluição colorimétrico para detecção da resistência à polimixina B em isolados clínicos de bacilos Gram-negativos. Foram testadas 11 isolados clínicos, sendo 8 E. coli e 3 K. pneumoniae. O método colorimétrico de microeluição da polimixina B foi uma adaptação do método da eluição da colistina, adicionando resazurina no final do processo. Os resultados foram interpretados de acordo com os pontos de corte estabelecidos para polimixina B pelo BrCAST. Quando avaliado os resultados da microeluição com e sem a resazurina, 7 isolados estavam em concordância categórica e 9 e 10 isolados, respectivamente, em concordância essencial com o método de referência. Diante do exposto, podemos concluir que tanto o teste da microeluição como o teste colorimétrico de microeluição da polimixina B demonstraram facilidade de realização, além de requerer materiais comumente encontrados em laboratórios de microbiologia de rotina. Ademais, um método envolvendo a polimixina B, a qual é a polimixina utilizada no Brasil, vem fomentar o estudo de novos testes de suscetibilidade frente às polimixinas.4SimBrasilFCF - Faculdade de Ciências FarmacêuticasManaus-AMFarmácia - Bacharelado - ManausMagalhães, Karen Regina Carim da Costahttp://lattes.cnpq.br/4015149205638769Dalmolin, Tanise Vendruscolohttp://lattes.cnpq.br/3986624670455667Azevedo, Maria Ermelinda Filgueiras dehttp://lattes.cnpq.br/1763660104203876Silva, Fabiana Brandão Alveshttp://lattes.cnpq.br/1431427028384548https://orcid.org/0000-0001-9975-6470https://orcid.org/0000-0002-3846-4367https://orcid.org/0000-0002-3846-4367https://orcid.org/0000-0001-8358-8062Fernandes, Tássia Péres2022-05-23T17:34:07Z2022-05-312022-05-23T17:34:07Z2022-04-27info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bachelorThesishttp://riu.ufam.edu.br/handle/prefix/6237porBAKTHAVATCHALAM, Yamuna Devi; PRAGASAM, Agila Kumari; BISWAS, Indranil; VEERARAGHAVAN, Balaji. Polymyxin susceptibility testing, interpretative breakpoints and resistance mechanisms: an update. Journal Of Global Antimicrobial Resistance, v. 12, p. 124-136, mar. 2018. http://dx.doi.org/10.1016/j.jgar.2017.09.011. BrCAST. Tabelas de pontos de corte para interpretação de CIMs e diâmetros de halos. Brazilian Committee on Antimicrobial Susceptibility Testing. Versão 2.0. 2021. CIELO, Naiany C. et al. Polymyxin B broth disk elution: a feasible and accurate methodology to determine polymyxin b susceptibility in Enterobacterales. Diagnostic Microbiology And Infectious Disease, v. 98, n. 2, p. 115099, out. 2020. http://dx.doi.org/10.1016/j.diagmicrobio.2020.115099. DALMOLIN, Tanise Vendruscolo; et al. Elution methods to evaluate colistin susceptibility of Gram-negative rods. Diagnostic Microbiology And Infectious Disease, v. 96, n. 1, p. 114910, jan. 2020. http://dx.doi.org/10.1016/j.diagmicrobio.2019.114910. DALMOLIN, Tanise Vendruscolo; LIMA-MORALES, Daiana; BARTH, Afonso Luis. Plasmid-mediated Colistin Resistance: What Do We Know? Journal of Infectiology, v. 1, n. 2, p. 16-22, 2018. DELIBERALI, Bruno; et al. Prevalência de bacilos Gram-negativos não fermentadores de pacientes internados em Porto Alegre-RS. Jornal Brasileiro de Patologia e Medicina Laboratorial, v. 47, n. 5, p. 529-534, out. 2011. http://dx.doi.org/10.1590/s1676-24442011000500006. EUCAST. Colistin. Rationale for the EUCAST clinical breakpoints, version 1.0. n. October, p. 1–14, 2010. EZADI, Fereshteh; ARDEBILI, Abdollah; MIRNEJAD, Reza. Antimicrobial Susceptibility Testing for Polymyxins: challenges, issues, and recommendations. Journal Of Clinical Microbiology, v. 57, n. 4, p. 1-20, abr. 2019. http://dx.doi.org/10.1128/jcm.01390-18. GALES, A. C.; JONES, R. N.; SADER, H. S. Contemporary activity of colistin and polymyxin B against a worldwide collection of Gram-negative pathogens: results from the sentry antimicrobial surveillance program (2006-09). Journal Of Antimicrobial Chemotherapy, v. 66, n. 9, p. 2070-2074, 29 jun. 2011. http://dx.doi.org/10.1093/jac/dkr239. GARG, Suneel Kumar; et al. Resurgence of Polymyxin B for MDR/XDR GramNegative Infections: an overview of current evidence. Critical Care Research And Practice, v. 2017, p. 1-10, 2017. http://dx.doi.org/10.1155/2017/3635609. 24 HUMPHRIES, Romney M.; et al. Multicenter Evaluation of Colistin Broth Disk Elution and Colistin Agar Test: a report from the clinical and laboratory standards institute. Journal Of Clinical Microbiology, v. 57, n. 11, p. 1-10, nov. 2019. http://dx.doi.org/10.1128/jcm.01269-19. KVITKO, Carlos Henrique Cezimbra. Eficácia da polimixina B no tratamento de bacteremias por Pseudomonas aeruginosa. 2010. 80 f. Dissertação (Mestrado) - Curso de Programa de Pós Graduação em Medicina, Universidade Federal do Rio Grande do Sul, Porto Alegre, 2010. LIU, Yi-Yun; et al. Emergence of plasmid-mediated colistin resistance mechanism MCR-1 in animals and human beings in China: a microbiological and molecular biological study. The Lancet Infectious Diseases, v. 16, n. 2, p. 161- 168, fev. 2016. http://dx.doi.org/10.1016/s1473-3099(15)00424-7. MOUBARECK, Carole Ayoub. Polymyxins and Bacterial Membranes: a review of antibacterial activity and mechanisms of resistance. Membranes, v. 10, n. 8, p. 181, 8 ago. 2020. http://dx.doi.org/10.3390/membranes10080181. POIREL, Laurent; JAYOL, Aurélie; NORDMANN, Patrice. Polymyxins: antibacterial activity, susceptibility testing, and resistance mechanisms encoded by plasmids or chromosomes. Clinical Microbiology Reviews, v. 30, n. 2, p. 557-596, abr. 2017. http://dx.doi.org/10.1128/cmr.00064-16. SADER, Helio S.; FARRELL, David J.; JONES, Ronald N. Susceptibility of Klebsiella spp. to colistin and polymyxin B: results from the sentry antimicrobial surveillance program (2006-2009). International Journal Of Antimicrobial Agents, v. 37, n. 2, p. 174-175, fev. 2011. http://dx.doi.org/10.1016/j.ijantimicag.2010.10.005. SARKER, Satyajit D.; NAHAR, Lutfun; KUMARASAMY, Yashodharan. Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods, v. 42, n. 4, p. 321-324, ago. 2007. http://dx.doi.org/10.1016/j.ymeth.2007.01.006. SHARAFI, Toktam; ARDEBILI, Abdollah. Plastic binding feature of polymyxins: the effect on mic susceptibility measurements. Infection And Drug Resistance, v. 12, p. 2649-2653, ago. 2019. http://dx.doi.org/10.2147/idr.s219130. SIMNER, Patricia J.; et al. Two-Site Evaluation of the Colistin Broth Disk Elution Test To Determine Colistin In Vitro Activity against Gram-Negative Bacilli. Journal Of Clinical Microbiology, v. 57, n. 2, p. 1-7, fev. 2019. http://dx.doi.org/10.1128/jcm.01163-18info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFAMinstname:Universidade Federal do Amazonas (UFAM)instacron:UFAM2022-05-23T17:35:10Zoai:localhost:prefix/6237Repositório InstitucionalPUBhttp://riu.ufam.edu.br/oai/requestopendoar:2022-05-23T17:35:10Repositório Institucional da UFAM - Universidade Federal do Amazonas (UFAM)false
dc.title.none.fl_str_mv Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
title Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
spellingShingle Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
Fernandes, Tássia Péres
Enterobacteriaceae
Polimixina B.
Polimixinas
Resistência bacteriana a antibióticos
CIÊNCIAS DA SAÚDE
Resazurina
Calorimetria de microeluição
Propriedade policatiônica
Microeluição da colistina em placa
Acinetobacter spp.
title_short Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
title_full Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
title_fullStr Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
title_full_unstemmed Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
title_sort Método de microeluição colorimétrico: detecção da resistência à polimixina B em bacilos Gram-negativos
author Fernandes, Tássia Péres
author_facet Fernandes, Tássia Péres
author_role author
dc.contributor.none.fl_str_mv Magalhães, Karen Regina Carim da Costa
http://lattes.cnpq.br/4015149205638769
Dalmolin, Tanise Vendruscolo
http://lattes.cnpq.br/3986624670455667
Azevedo, Maria Ermelinda Filgueiras de
http://lattes.cnpq.br/1763660104203876
Silva, Fabiana Brandão Alves
http://lattes.cnpq.br/1431427028384548
https://orcid.org/0000-0001-9975-6470
https://orcid.org/0000-0002-3846-4367
https://orcid.org/0000-0002-3846-4367
https://orcid.org/0000-0001-8358-8062
dc.contributor.author.fl_str_mv Fernandes, Tássia Péres
dc.subject.por.fl_str_mv Enterobacteriaceae
Polimixina B.
Polimixinas
Resistência bacteriana a antibióticos
CIÊNCIAS DA SAÚDE
Resazurina
Calorimetria de microeluição
Propriedade policatiônica
Microeluição da colistina em placa
Acinetobacter spp.
topic Enterobacteriaceae
Polimixina B.
Polimixinas
Resistência bacteriana a antibióticos
CIÊNCIAS DA SAÚDE
Resazurina
Calorimetria de microeluição
Propriedade policatiônica
Microeluição da colistina em placa
Acinetobacter spp.
description In the last years, the reports of polymyxins resistance have increased considerably, so efficient, reliable, fast and easily interpreted testing to determine polymyxins susceptibility is necessary. The present study aimed to develop and evaluate the performance of a colorimetric microelution method for detecting polymyxin B resistance in clinical isolates of Gram-negative bacilli. The tests were performed with 11 clinical isolates, including 8 E. coli and 3 K. pneumoniae. The colorimetric polymyxin B microelution method was an adaptation of the colistin broth disk elution test, adding resazurin at the end of the process. The results were interpreted using the breakpoints of the BrCAST. When evaluating the microelution results with and without resazurin, 7 isolates presented categorical agreement and 9 and 10 isolates, respectively, presented essential agreement with the reference method. In conclusion, both polymyxin B microelution test and the colorimetric polymyxin B microelution test were easy to perform, requiring materials commonly found in routine microbiology laboratories. Furthermore, a method involving polymyxin B, which is the polymyxin used in Brazil, encourages the study of new susceptibility tests against polymyxins.
publishDate 2022
dc.date.none.fl_str_mv 2022-05-23T17:34:07Z
2022-05-31
2022-05-23T17:34:07Z
2022-04-27
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/bachelorThesis
format bachelorThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://riu.ufam.edu.br/handle/prefix/6237
url http://riu.ufam.edu.br/handle/prefix/6237
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv BAKTHAVATCHALAM, Yamuna Devi; PRAGASAM, Agila Kumari; BISWAS, Indranil; VEERARAGHAVAN, Balaji. Polymyxin susceptibility testing, interpretative breakpoints and resistance mechanisms: an update. Journal Of Global Antimicrobial Resistance, v. 12, p. 124-136, mar. 2018. http://dx.doi.org/10.1016/j.jgar.2017.09.011. BrCAST. Tabelas de pontos de corte para interpretação de CIMs e diâmetros de halos. Brazilian Committee on Antimicrobial Susceptibility Testing. Versão 2.0. 2021. CIELO, Naiany C. et al. Polymyxin B broth disk elution: a feasible and accurate methodology to determine polymyxin b susceptibility in Enterobacterales. Diagnostic Microbiology And Infectious Disease, v. 98, n. 2, p. 115099, out. 2020. http://dx.doi.org/10.1016/j.diagmicrobio.2020.115099. DALMOLIN, Tanise Vendruscolo; et al. Elution methods to evaluate colistin susceptibility of Gram-negative rods. Diagnostic Microbiology And Infectious Disease, v. 96, n. 1, p. 114910, jan. 2020. http://dx.doi.org/10.1016/j.diagmicrobio.2019.114910. DALMOLIN, Tanise Vendruscolo; LIMA-MORALES, Daiana; BARTH, Afonso Luis. Plasmid-mediated Colistin Resistance: What Do We Know? Journal of Infectiology, v. 1, n. 2, p. 16-22, 2018. DELIBERALI, Bruno; et al. Prevalência de bacilos Gram-negativos não fermentadores de pacientes internados em Porto Alegre-RS. Jornal Brasileiro de Patologia e Medicina Laboratorial, v. 47, n. 5, p. 529-534, out. 2011. http://dx.doi.org/10.1590/s1676-24442011000500006. EUCAST. Colistin. Rationale for the EUCAST clinical breakpoints, version 1.0. n. October, p. 1–14, 2010. EZADI, Fereshteh; ARDEBILI, Abdollah; MIRNEJAD, Reza. Antimicrobial Susceptibility Testing for Polymyxins: challenges, issues, and recommendations. Journal Of Clinical Microbiology, v. 57, n. 4, p. 1-20, abr. 2019. http://dx.doi.org/10.1128/jcm.01390-18. GALES, A. C.; JONES, R. N.; SADER, H. S. Contemporary activity of colistin and polymyxin B against a worldwide collection of Gram-negative pathogens: results from the sentry antimicrobial surveillance program (2006-09). Journal Of Antimicrobial Chemotherapy, v. 66, n. 9, p. 2070-2074, 29 jun. 2011. http://dx.doi.org/10.1093/jac/dkr239. GARG, Suneel Kumar; et al. Resurgence of Polymyxin B for MDR/XDR GramNegative Infections: an overview of current evidence. Critical Care Research And Practice, v. 2017, p. 1-10, 2017. http://dx.doi.org/10.1155/2017/3635609. 24 HUMPHRIES, Romney M.; et al. Multicenter Evaluation of Colistin Broth Disk Elution and Colistin Agar Test: a report from the clinical and laboratory standards institute. Journal Of Clinical Microbiology, v. 57, n. 11, p. 1-10, nov. 2019. http://dx.doi.org/10.1128/jcm.01269-19. KVITKO, Carlos Henrique Cezimbra. Eficácia da polimixina B no tratamento de bacteremias por Pseudomonas aeruginosa. 2010. 80 f. Dissertação (Mestrado) - Curso de Programa de Pós Graduação em Medicina, Universidade Federal do Rio Grande do Sul, Porto Alegre, 2010. LIU, Yi-Yun; et al. Emergence of plasmid-mediated colistin resistance mechanism MCR-1 in animals and human beings in China: a microbiological and molecular biological study. The Lancet Infectious Diseases, v. 16, n. 2, p. 161- 168, fev. 2016. http://dx.doi.org/10.1016/s1473-3099(15)00424-7. MOUBARECK, Carole Ayoub. Polymyxins and Bacterial Membranes: a review of antibacterial activity and mechanisms of resistance. Membranes, v. 10, n. 8, p. 181, 8 ago. 2020. http://dx.doi.org/10.3390/membranes10080181. POIREL, Laurent; JAYOL, Aurélie; NORDMANN, Patrice. Polymyxins: antibacterial activity, susceptibility testing, and resistance mechanisms encoded by plasmids or chromosomes. Clinical Microbiology Reviews, v. 30, n. 2, p. 557-596, abr. 2017. http://dx.doi.org/10.1128/cmr.00064-16. SADER, Helio S.; FARRELL, David J.; JONES, Ronald N. Susceptibility of Klebsiella spp. to colistin and polymyxin B: results from the sentry antimicrobial surveillance program (2006-2009). International Journal Of Antimicrobial Agents, v. 37, n. 2, p. 174-175, fev. 2011. http://dx.doi.org/10.1016/j.ijantimicag.2010.10.005. SARKER, Satyajit D.; NAHAR, Lutfun; KUMARASAMY, Yashodharan. Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods, v. 42, n. 4, p. 321-324, ago. 2007. http://dx.doi.org/10.1016/j.ymeth.2007.01.006. SHARAFI, Toktam; ARDEBILI, Abdollah. Plastic binding feature of polymyxins: the effect on mic susceptibility measurements. Infection And Drug Resistance, v. 12, p. 2649-2653, ago. 2019. http://dx.doi.org/10.2147/idr.s219130. SIMNER, Patricia J.; et al. Two-Site Evaluation of the Colistin Broth Disk Elution Test To Determine Colistin In Vitro Activity against Gram-Negative Bacilli. Journal Of Clinical Microbiology, v. 57, n. 2, p. 1-7, fev. 2019. http://dx.doi.org/10.1128/jcm.01163-18
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Brasil
FCF - Faculdade de Ciências Farmacêuticas
Manaus-AM
Farmácia - Bacharelado - Manaus
publisher.none.fl_str_mv Brasil
FCF - Faculdade de Ciências Farmacêuticas
Manaus-AM
Farmácia - Bacharelado - Manaus
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFAM
instname:Universidade Federal do Amazonas (UFAM)
instacron:UFAM
instname_str Universidade Federal do Amazonas (UFAM)
instacron_str UFAM
institution UFAM
reponame_str Repositório Institucional da UFAM
collection Repositório Institucional da UFAM
repository.name.fl_str_mv Repositório Institucional da UFAM - Universidade Federal do Amazonas (UFAM)
repository.mail.fl_str_mv
_version_ 1813274314786471936