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Screening and in silico characterization of prophages in Helicobacter pylori clinical strains

datacite.subject.fosCiências Naturais
dc.contributor.authorFerreira, Rute
dc.contributor.authorPinto, Graça
dc.contributor.authorPresa, Eva
dc.contributor.authorOleastro, Mónica
dc.contributor.authorSilva, Catarina
dc.contributor.authorVieira, Luís
dc.contributor.authorSousa, Claúdia
dc.contributor.authorPires, Diana
dc.contributor.authorFigueiredo, Ceu
dc.contributor.authorMelo, Luís
dc.date.accessioned2025-04-02T13:21:11Z
dc.date.available2025-04-02T13:21:11Z
dc.date.issued2024-10-03
dc.description.abstractThe increase of antibiotic resistance calls for alternatives to control Helicobacter pylori, a Gram-negative bacterium associated with various gastric diseases. Bacteriophages (phages) can be highly effective in the treatment of pathogenic bacteria. Here, we developed a method to identify prophages in H. pylori genomes aiming at their future use in therapy. A polymerase chain reaction (PCR)-based technique tested five primer pairs on 74 clinical H. pylori strains. After the PCR screening, 14 strains most likely to carry prophages were fully sequenced. After that, a more holistic approach was taken by studying the complete genome of the strains. This study allowed us to identify 12 intact prophage sequences, which were then characterized concerning their morphology, virulence, and antibiotic-resistance genes. To understand the variability of prophages, a phylogenetic analysis using the sequences of all H. pylori phages reported to date was performed. Overall, we increased the efficiency of identifying complete prophages to 54.1 %. Genes with homology to potential virulence factors were identified in some new prophages. Phylogenetic analysis revealed a close relationship among H. pylori-phages, although there are phages with different geographical origins. This study provides a deeper understanding of H. pylori-phages, providing valuable insights into their potential use in therapy.eng
dc.description.abstractQuestions answered in this article: - What were the findings regarding the number of prophages identified in the H. pylori genomes? The study identified twelve new prophages in the genomes of Helicobacter pylori, with at least 10 out of the 14 sequenced strains containing at least one intact prophage, indicating a significant presence of these genetic elements in the clinical strains examined; -What percentage of H. pylori strains are compatible with the presence of prophage genes? The analysis of Helicobacter pylori genomes indicates that approximately 20% of strains are compatible with the presence of prophage genes; - What Polymerase Chain Reaction (PCR) techniques were employed to screen for prophages in H. pylori strains? The Polymerase Chain Reaction (PCR) techniques employed to screen for prophages in Helicobacter pylori strains included the use of four different primer sets targeting two regions of the integrase gene, the holin gene, and the insertion site region between specific genes. This approach aimed to enhance the detection of prophages in the clinical strains; What morphological characteristics are commonly associated with H. pylori prophages? H. pylori prophages are commonly associated with a podovirus-like morphology, as indicated by the results obtained with VIRFAM. This morphological characteristic aligns with most reported H. pylori prophages, which exhibit this specific structure; - What challenges exist in the genomic analysis and characterization of H. pylori prophages? The genomic analysis and characterization of Helicobacter pylori prophages face challenges such as the alarming rates of antibiotic resistance, which complicate treatment approaches and necessitate alternative strategies. Additionally, the variability in the genomic size and GC content among different H. pylori strains may pose difficulties in standardizing genomic analyses.por
dc.description.sponsorshipThis work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit, and by LABBELS – Associate Laboratory in Biotechnology, Bioengineering, and Microelectromechanical Systems, LA/P/0029/2020 and Project Helicophage PTDC/SAU-PUB/29182/2017 [POCI-01-0145-FEDER-029182]. Rute Ferreira acknowledges the FCT grant (https://doi.org/10.54499/SFRH/BD/146496/2019). Luís D. R. Melo acknowledges funding from the FCT through the Scientific Employment Stimulus Program (https://doi.org/10.54499/2021.00221.CEECIND/CP1664/CT0002).
dc.identifier.citationMicrobes Infect. 2024 Oct 3:105429. doi: 10.1016/j.micinf.2024.105429. Online ahead of print
dc.identifier.doi10.1016/j.micinf.2024.105429
dc.identifier.issn1286-4579
dc.identifier.pmid39368610
dc.identifier.urihttp://hdl.handle.net/10400.18/10466
dc.language.isoeng
dc.peerreviewedyes
dc.publisherElsevier
dc.relationPTDC/SAU-PUB/29182/2017
dc.relationCentre of Biological Engineering of the University of Minho
dc.relationA new strategy to combat Helicobacter pylori infections based on chimeric phages
dc.relation.hasversionhttps://linkinghub.elsevier.com/retrieve/pii/S1286457924001710
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectHelicobacter pylori
dc.subjectAntimicrobial Resistance Genes
dc.subjectPCR
dc.subjectProphages
dc.subjectVirulence Factors
dc.subjectInfecções Gastrointestinais
dc.subjectResistência aos Antimicrobianos
dc.titleScreening and in silico characterization of prophages in Helicobacter pylori clinical strainspor
dc.typeclinical study
dspace.entity.typePublication
oaire.awardTitleCentre of Biological Engineering of the University of Minho
oaire.awardTitleA new strategy to combat Helicobacter pylori infections based on chimeric phages
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/9471 - RIDTI/PTDC%2FSAU-PUB%2F29182%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04469%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/POR_NORTE/SFRH%2FBD%2F146496%2F2019/PT
oaire.citation.startPage105429
oaire.citation.titleMicrobes and Infection
oaire.fundingStream9471 - RIDTI
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStreamPOR_NORTE
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
relation.isProjectOfPublication4118b900-7c6f-4ebb-9ffb-f7f7b5db8b8a
relation.isProjectOfPublication7943bed3-8247-4296-abf0-957ef0a00120
relation.isProjectOfPublication05dc65c1-1b8d-4cdf-9ef4-30d761fd9761
relation.isProjectOfPublication.latestForDiscovery4118b900-7c6f-4ebb-9ffb-f7f7b5db8b8a

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