Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates
Autor(a) principal: | |
---|---|
Data de Publicação: | 2022 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10773/34469 |
Resumo: | Haemophilus influenzae is a human pathogen responsible for severe invasive disease such as meningitis, septicaemia, epiglottitis and pneumonia in children and adults. Beta-lactams antibiotics, such as ampicillin, are widely used as empirical treatment of these infections. The emergence of resistant strains, as a result of selective antibiotic pressure, has become an issue of concern. The aim of this study was to characterize the collection of 309 Haemophilus influenzae invasive isolates received between 2015 and 2021, in Haemophilus influenzae Reference Laboratory, from National Institute of Health Dr Ricardo Jorge, in Lisbon. Molecular characterization of ampicillin resistance mechanisms and genetic diversity was investigated to follow changes in epidemiology of Haemophilus influenzae invasive disease. Antimicrobial susceptibility was determined for 298 viable isolates, against 12 antibiotics. The ftsI gene, encoding the transpeptidase domain of penicillinbinding- protein 3, was sequenced for 74 isolates, both β-lactamase-producers and non-producers. Multilocus sequence typing was performed to examine genetic relatedness among 277 isolates. Previous characterization of capsular type showed that most isolates were nonencapsulated (72.8%) and 27.2% were encapsulated, with 12.3% serotype b, 7.8% serotype a, 4.8% serotype f and 2.3% serotype e. Overall, most isolates were susceptible to the antibiotics studied, with exception of 16.1% resistant to ampicillin and 21.1% resistant to trimethoprim-sulfamethoxazole. β-lactamaseproducers accounted for 13.3% of the isolates. Sequence analysis of ftsI gene, showed mutations in 70.3% of the isolates (52/74). Of the 74 isolates analysed, 52.7% were classified as genotypically β- lactamase-negative ampicillin-resistant and 8.1% as genotypically β-lactamasepositive ampicillin-clavulanic acid-resistant, according to their mutational patterns. Multilocus sequence typing revealed high genetic diversity among nonencapsulated isolates, while encapsulated isolates were clonal within each serotype. Two sequence types were predominant among nonencapsulated isolates, ST103 and ST1034, being associated, respectively, with β-lactamaseproducers and non-β-lactamase-producers. Our data reiterate the need for continued surveillance of resistance in Haemophilus influenzae infections, which will help to develop Public Health prevention strategies. |
id |
RCAP_4c7e5fd9781fe3f648752dd2fbaa11af |
---|---|
oai_identifier_str |
oai:ria.ua.pt:10773/34469 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolatesHaemophilus influenzaeβ-lactam resistancePenicillin binding proteins (PBP3)ftsI geneGenetic relatednessMultilocus-sequence-typingHaemophilus influenzae is a human pathogen responsible for severe invasive disease such as meningitis, septicaemia, epiglottitis and pneumonia in children and adults. Beta-lactams antibiotics, such as ampicillin, are widely used as empirical treatment of these infections. The emergence of resistant strains, as a result of selective antibiotic pressure, has become an issue of concern. The aim of this study was to characterize the collection of 309 Haemophilus influenzae invasive isolates received between 2015 and 2021, in Haemophilus influenzae Reference Laboratory, from National Institute of Health Dr Ricardo Jorge, in Lisbon. Molecular characterization of ampicillin resistance mechanisms and genetic diversity was investigated to follow changes in epidemiology of Haemophilus influenzae invasive disease. Antimicrobial susceptibility was determined for 298 viable isolates, against 12 antibiotics. The ftsI gene, encoding the transpeptidase domain of penicillinbinding- protein 3, was sequenced for 74 isolates, both β-lactamase-producers and non-producers. Multilocus sequence typing was performed to examine genetic relatedness among 277 isolates. Previous characterization of capsular type showed that most isolates were nonencapsulated (72.8%) and 27.2% were encapsulated, with 12.3% serotype b, 7.8% serotype a, 4.8% serotype f and 2.3% serotype e. Overall, most isolates were susceptible to the antibiotics studied, with exception of 16.1% resistant to ampicillin and 21.1% resistant to trimethoprim-sulfamethoxazole. β-lactamaseproducers accounted for 13.3% of the isolates. Sequence analysis of ftsI gene, showed mutations in 70.3% of the isolates (52/74). Of the 74 isolates analysed, 52.7% were classified as genotypically β- lactamase-negative ampicillin-resistant and 8.1% as genotypically β-lactamasepositive ampicillin-clavulanic acid-resistant, according to their mutational patterns. Multilocus sequence typing revealed high genetic diversity among nonencapsulated isolates, while encapsulated isolates were clonal within each serotype. Two sequence types were predominant among nonencapsulated isolates, ST103 and ST1034, being associated, respectively, with β-lactamaseproducers and non-β-lactamase-producers. Our data reiterate the need for continued surveillance of resistance in Haemophilus influenzae infections, which will help to develop Public Health prevention strategies.Haemophilus influenzae é um agente patogénico humano, responsável por doenças invasivas graves em crianças e adultos, como meningite, septicemia, epiglotite e pneumonia. Os antibióticos beta-lactâmicos, como a ampicilina, são os mais utilizados no tratamento empírico destas infecções. O aparecimento de estirpes resistentes, resultado da pressão antibiótica selectiva, tornou-se uma preocupação em saúde pública. O objectivo deste estudo foi caracterizar a colecção de 309 isolados invasivos de Haemophilus influenzae, recebidos entre 2015 e 2021, no Laboratório de Referência de Haemophilus influenzae, do Instituto Nacional de Saúde Dr Ricardo Jorge, sediado em Lisboa. A caracterização molecular dos mecanismos de resistência à ampicilina e a diversidade genética foram investigadas para monitorizar alterações na epidemiologia da doença invasiva de Haemophilus influenzae. A susceptibilidade antimicrobiana foi determinada para 12 antibióticos, em 298 isolados viáveis. O gene ftsI, que codifica o domínio transpeptidase da proteína de ligação à penicilina 3, foi sequenciado em 74 isolados, produtores e não produtores de β-lactamases. A técnica de multilocus sequence typing foi realizada para analisar a relação genética entre 277 isolados. A caracterização do tipo capsular, previamente determinada, mostrou que a maioria dos isolados eram não capsulados (72,8%), sendo os restantes 27,2% capsulados: 12,3% serotipo b, 7,8% serotipo a, 4,8% serotipo f e 2,3% serotipo e. No geral, a maioria dos isolados eram susceptíveis aos antibióticos estudados, com excepção de 16,1% resistentes à ampicilina e 21,1% resistentes ao trimetoprim-sulfametoxazol. A produção de β- lactamases foi detetada em 13,3% dos isolados. A análise das sequências do gene ftsI revelou que 70,3% dos isolados (52/74) tinham mutações. Dos 74 isolados analisados, 52,7% foram classificados como genotipicamente β-lactamase-negativos resistentes à ampicilina e 8,1% como genotipicamente β-lactamase-positivos resistentes à ampicilina-ácido clavulânico, de acordo com os seus padrões mutacionais. A técnica de multilocus sequence typing demonstrou uma elevada diversidade genética entre os isolados não capsulados, enquanto que os isolados capsulados eram clonais dentro de cada serotipo. Dois sequence types revelaram-se predominantes entre os isolados não capsulados, ST103 e ST1034, estando associados, respectivamente, a isolados β-lactamase-positivos e β-lactamase-negativos. Os nossos dados reiteraram a necessidade da vigilância contínua das infecções por Haemophilus influenzae, que ajudarão a definir tendências na resistência à ampicilina, bem como a prevenir a propagação de estirpes resistentes, através de medidas terapêuticas e preventivas adequadas.2024-07-27T00:00:00Z2022-07-14T00:00:00Z2022-07-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/34469engFrederico, Andreia Filipa Martinsinfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T12:06:41Zoai:ria.ua.pt:10773/34469Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:05:49.601903Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates |
title |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates |
spellingShingle |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates Frederico, Andreia Filipa Martins Haemophilus influenzae β-lactam resistance Penicillin binding proteins (PBP3) ftsI gene Genetic relatedness Multilocus-sequence-typing |
title_short |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates |
title_full |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates |
title_fullStr |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates |
title_full_unstemmed |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates |
title_sort |
Beta-lactam resistance pattern and sequence type in Portuguese Haemophilus influenzae invasive isolates |
author |
Frederico, Andreia Filipa Martins |
author_facet |
Frederico, Andreia Filipa Martins |
author_role |
author |
dc.contributor.author.fl_str_mv |
Frederico, Andreia Filipa Martins |
dc.subject.por.fl_str_mv |
Haemophilus influenzae β-lactam resistance Penicillin binding proteins (PBP3) ftsI gene Genetic relatedness Multilocus-sequence-typing |
topic |
Haemophilus influenzae β-lactam resistance Penicillin binding proteins (PBP3) ftsI gene Genetic relatedness Multilocus-sequence-typing |
description |
Haemophilus influenzae is a human pathogen responsible for severe invasive disease such as meningitis, septicaemia, epiglottitis and pneumonia in children and adults. Beta-lactams antibiotics, such as ampicillin, are widely used as empirical treatment of these infections. The emergence of resistant strains, as a result of selective antibiotic pressure, has become an issue of concern. The aim of this study was to characterize the collection of 309 Haemophilus influenzae invasive isolates received between 2015 and 2021, in Haemophilus influenzae Reference Laboratory, from National Institute of Health Dr Ricardo Jorge, in Lisbon. Molecular characterization of ampicillin resistance mechanisms and genetic diversity was investigated to follow changes in epidemiology of Haemophilus influenzae invasive disease. Antimicrobial susceptibility was determined for 298 viable isolates, against 12 antibiotics. The ftsI gene, encoding the transpeptidase domain of penicillinbinding- protein 3, was sequenced for 74 isolates, both β-lactamase-producers and non-producers. Multilocus sequence typing was performed to examine genetic relatedness among 277 isolates. Previous characterization of capsular type showed that most isolates were nonencapsulated (72.8%) and 27.2% were encapsulated, with 12.3% serotype b, 7.8% serotype a, 4.8% serotype f and 2.3% serotype e. Overall, most isolates were susceptible to the antibiotics studied, with exception of 16.1% resistant to ampicillin and 21.1% resistant to trimethoprim-sulfamethoxazole. β-lactamaseproducers accounted for 13.3% of the isolates. Sequence analysis of ftsI gene, showed mutations in 70.3% of the isolates (52/74). Of the 74 isolates analysed, 52.7% were classified as genotypically β- lactamase-negative ampicillin-resistant and 8.1% as genotypically β-lactamasepositive ampicillin-clavulanic acid-resistant, according to their mutational patterns. Multilocus sequence typing revealed high genetic diversity among nonencapsulated isolates, while encapsulated isolates were clonal within each serotype. Two sequence types were predominant among nonencapsulated isolates, ST103 and ST1034, being associated, respectively, with β-lactamaseproducers and non-β-lactamase-producers. Our data reiterate the need for continued surveillance of resistance in Haemophilus influenzae infections, which will help to develop Public Health prevention strategies. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-07-14T00:00:00Z 2022-07-14 2024-07-27T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10773/34469 |
url |
http://hdl.handle.net/10773/34469 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
_version_ |
1799137713339760640 |