Pyrosequencing analysis for rapid and accurate detection of clarithromycin resistance-associated mutations in Iranian Helicobacter pylori isolates.
Humans
Clarithromycin
/ pharmacology
Helicobacter pylori
/ genetics
Anti-Bacterial Agents
/ pharmacology
Iran
Drug Resistance, Bacterial
/ genetics
Polymerase Chain Reaction
/ methods
Mutation
Helicobacter Infections
/ drug therapy
Microbial Sensitivity Tests
RNA, Ribosomal, 23S
/ genetics
High-Throughput Nucleotide Sequencing
Antibiotic resistance
Clarithromycin
Gene mutation
Helicobacter pylori
Pyrosequencing
Sanger sequencing
Journal
BMC research notes
ISSN: 1756-0500
Titre abrégé: BMC Res Notes
Pays: England
ID NLM: 101462768
Informations de publication
Date de publication:
06 Jul 2023
06 Jul 2023
Historique:
received:
18
03
2023
accepted:
27
06
2023
medline:
10
7
2023
pubmed:
7
7
2023
entrez:
6
7
2023
Statut:
epublish
Résumé
Treatment of Helicobacter pylori (H. pylori) infection has become challenging following the development of primary antibiotic resistance. A primary therapeutic regimen for H. pylori eradication includes clarithromycin; however, the presence of point mutations within the 23S rRNA sequence of H. pylori contributes to clarithromycin resistance and eradication failure. Thus, we aimed to develop a rapid and precise method to determine clarithromycin resistance-related point mutations using the pyrosequencing method. H. pylori was isolated from 82 gastric biopsy samples and minimal inhibitory concentration (MIC) was evaluated using the agar dilution method. Clarithromycin resistance-associated point mutations were detected by Sanger sequencing, from which 11 isolates were chosen for pyrosequencing. Our results demonstrated a 43.9% (36/82) prevalence in resistance to clarithromycin. The A2143G mutation was detected in 8.3% (4/48) of H. pylori isolates followed by A2142G (6.2%), C2195T (4.1%), T2182C (4.1%), and C2288T (2%). Although the C2195T mutation was only detected by Sanger sequencing, the overall results from pyrosequencing and Sanger sequencing platforms were comparable. Pyrosequencing could be used as a rapid and practical platform in clinical laboratories to determine the susceptibility profile of H. pylori isolates. This might pave the way for efficient H. pylori eradication upon detection.
Sections du résumé
BACKGROUND
BACKGROUND
Treatment of Helicobacter pylori (H. pylori) infection has become challenging following the development of primary antibiotic resistance. A primary therapeutic regimen for H. pylori eradication includes clarithromycin; however, the presence of point mutations within the 23S rRNA sequence of H. pylori contributes to clarithromycin resistance and eradication failure. Thus, we aimed to develop a rapid and precise method to determine clarithromycin resistance-related point mutations using the pyrosequencing method.
METHODS AND RESULTS
RESULTS
H. pylori was isolated from 82 gastric biopsy samples and minimal inhibitory concentration (MIC) was evaluated using the agar dilution method. Clarithromycin resistance-associated point mutations were detected by Sanger sequencing, from which 11 isolates were chosen for pyrosequencing. Our results demonstrated a 43.9% (36/82) prevalence in resistance to clarithromycin. The A2143G mutation was detected in 8.3% (4/48) of H. pylori isolates followed by A2142G (6.2%), C2195T (4.1%), T2182C (4.1%), and C2288T (2%). Although the C2195T mutation was only detected by Sanger sequencing, the overall results from pyrosequencing and Sanger sequencing platforms were comparable.
CONCLUSIONS
CONCLUSIONS
Pyrosequencing could be used as a rapid and practical platform in clinical laboratories to determine the susceptibility profile of H. pylori isolates. This might pave the way for efficient H. pylori eradication upon detection.
Identifiants
pubmed: 37415212
doi: 10.1186/s13104-023-06420-0
pii: 10.1186/s13104-023-06420-0
pmc: PMC10324197
doi:
Substances chimiques
Clarithromycin
H1250JIK0A
Anti-Bacterial Agents
0
RNA, Ribosomal, 23S
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
136Subventions
Organisme : Shahid Beheshti University of Medical Sciences
ID : RIGLD 878
Informations de copyright
© 2023. The Author(s).
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