1887

Journal of Medical Microbiology logo

Volume 73, Issue 5

Genetic variations of penicillin-binding protein 1A: insights into the current status of amoxicillin-based regimens for eradication in Malaysia No Access

Abstract

Resistance towards amoxicillin in causes significant therapeutic impasse in healthcare settings worldwide. In Malaysia, the standard treatment regimen includes a 14-day course of high-dose proton-pump inhibitor (rabeprazole, 20 mg) with amoxicillin (1000 mg) dual therapy.

The high eradication rate with amoxicillin-based treatment could be attributed to the primary resistance rates of amoxicillin being relatively low at 0%, however, a low rate of secondary resistance has been documented in Malaysia recently.

This study aims to investigate the amino acid mutations and related genetic variants in PBP1A of , correlating with amoxicillin resistance in the Malaysian population.

The full-length gene was amplified via PCR from 50 genomic DNA extracted from gastric biopsy samples of -positive treatment-naïve Malaysian patients. The sequences were then compared with reference strain ATCC 26695 for mutation and variant detection. A phylogenetic analysis of 50 sequences along with 43 additional sequences from the NCBI database was performed. These additional sequences included both amoxicillin-resistant strains (=20) and amoxicillin-sensitive strains (=23).

There was a total of 21 variants of amino acids, with three of them located in or near the PBP-motif (SKN402-404). The percentages of these three variants are as follows: K403X, 2%; S405I, 2% and E406K, 16%. Based on the genetic markers identified, the resistance rate for amoxicillin in our sample remained at 0%. The phylogenetic examination suggested that might exhibit unique conserved sequences within the Malaysian context.

Overall, the molecular analysis of PBP1A supported the therapeutic superiority of amoxicillin-based regimens. Therefore, it is crucial to continue monitoring the amoxicillin resistance background of with a larger sample size to ensure the sustained effectiveness of amoxicillin-based treatments in Malaysia.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): amoxicillin resistance , full-length sequencing , Helicobacter pylori , PBP1A and phylogenetic analysis
Funding
This study was supported by the:
  • UM (Award IIRG029A/B/C-2019)
    • Principle Award Recipient: SuatMoi Puah
© 2024 The Authors
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001832
2024-05-07
2024-05-20
Loading full text...

Full text loading...

References

  1. Kageyama C, Sato M, Sakae H, Obayashi Y, Kawahara Y et al. Increase in antibiotic resistant Helicobacter pylori in a University Hospital in Japan. Infect Drug Resist 2019; 12:597–602 [View Article] [PubMed]
     [Google Scholar]
  2. Hashemi SJ, Sheikh AF, Goodarzi H, Yadyad MJ, Seyedian SS et al. Genetic basis for metronidazole and clarithromycin resistance in Helicobacter pylori strains isolated from patients with gastroduodenal disorders. Infect Drug Resist 2019; 12:535–543 [View Article] [PubMed]
     [Google Scholar]
  3. Lee YC, Dore MP, Graham DY. Diagnosis and treatment of Helicobacter pylori infection. Annu Rev Med 2022; 73:183–195 [View Article] [PubMed]
     [Google Scholar]
  4. Malfertheiner P, Megraud F, O’Morain CA, Gisbert JP, Kuipers EJ et al. Management of Helicobacter pylori infection-the maastricht V/florence consensus report. Gut 2017; 66:6–30 [View Article] [PubMed]
     [Google Scholar]
  5. Goh K-L, Lee YY, Leow AH-R, Ali RAR, Ho SH et al. A Malaysian consensus report on the diagnosis and treatment of Helicobacter pylori infection. JGH Open 2023; 7:261–271 [View Article] [PubMed]
     [Google Scholar]
  6. Gerrits MM, van Vliet AHM, Kuipers EJ, Kusters JG. Helicobacter pylori and antimicrobial resistance: molecular mechanisms and clinical implications. Lancet Infect Dis 2006; 6:699–709 [View Article] [PubMed]
     [Google Scholar]
  7. Savoldi A, Carrara E, Graham DY, Conti M, Tacconelli E. Prevalence of antibiotic resistance in Helicobacter pylori: a systematic review and meta-analysis in World Health Organization regions. Gastroenterology 2018; 155:1372–1382 [View Article] [PubMed]
     [Google Scholar]
  8. Boyanova L, Hadzhiyski P, Gergova R, Markovska R. Evolution of Helicobacter pylori resistance to antibiotics: a topic of increasing concern. Antibiotics (Basel) 2023; 12:332 [View Article] [PubMed]
     [Google Scholar]
  9. Wang D, Guo Q, Yuan Y, Gong Y. The antibiotic resistance of Helicobacter pylori to five antibiotics and influencing factors in an area of China with a high risk of gastric cancer. BMC Microbiol 2019; 19:152 [View Article] [PubMed]
     [Google Scholar]
  10. Sholeh M, Maleki F, Krutova M, Bavari S, Golmoradi R et al. The increasing antimicrobial resistance of Helicobacter pylori in Iran: a systematic review and meta-analysis. Helicobacter 2020; 25:e12730 [View Article] [PubMed]
     [Google Scholar]
  11. Tran TT, Nguyen AT, Quach DT, Pham DT-H, Cao NM et al. Emergence of amoxicillin resistance and identification of novel mutations of the pbp1A gene in Helicobacter pylori in Vietnam. BMC Microbiol 2022; 22:41 [View Article] [PubMed]
     [Google Scholar]
  12. Boyanova L, Kandilarov N, Hadzhiyski P, Gergova R, Gergova G et al. Increase in amoxicillin resistance in Helicobacter pylori from Bulgarian patients over 15 years. Diagn Microbiol Infect Dis 2022; 104:115746 [View Article] [PubMed]
     [Google Scholar]
  13. Ahmad N, Zakaria WR, Mohamed R. Analysis of antibiotic susceptibility patterns of Helicobacter pylori isolates from Malaysia. Helicobacter 2011; 16:47–51 [View Article] [PubMed]
     [Google Scholar]
  14. Alfizah H, Norazah A, Hamizah R, Ramelah M. Resistotype of Helicobacter pylori isolates: the impact on eradication outcome. J Med Microbiol 2014; 63:703–709 [View Article] [PubMed]
     [Google Scholar]
  15. Goh KL, Navaratnam P. High Helicobacter pylori resistance to metronidazole but zero or low resistance to clarithromycin, levofloxacin, and other antibiotics in Malaysia. Helicobacter 2011; 16:241–245 [View Article] [PubMed]
     [Google Scholar]
  16. Hanafiah A, Binmaeil H, Raja Ali RA, Mohamed Rose I, Lopes BS. Molecular characterization and prevalence of antibiotic resistance in Helicobacter pylori isolates in Kuala Lumpur, Malaysia. Infect Drug Resist 2019; 12:3051–3061 [View Article] [PubMed]
     [Google Scholar]
  17. Teh X, Khosravi Y, Lee WC, Leow AHR, Loke MF et al. Functional and molecular surveillance of Helicobacter pylori antibiotic resistance in Kuala Lumpur. PLoS One 2014; 9:e101481 [View Article] [PubMed]
     [Google Scholar]
  18. Sukri A, Hanafiah A, Yusoff H, Shamsul Nizam NA, Nameyrra Z et al. Multidrug-resistant Helicobacter pylori strains: a five-year surveillance study and its genome characteristics. Antibiotics 2022; 11:1391 [View Article] [PubMed]
     [Google Scholar]
  19. Attaran B, Salehi N, Ghadiri B, Esmaeili M, Kalateh S et al. The penicillin binding protein 1A of Helicobacter pylori, its amoxicillin binding site and access routes. Gut Pathog 2021; 13:43 [View Article] [PubMed]
     [Google Scholar]
  20. Caveney NA, Li FK, Strynadka NC. Enzyme structures of the bacterial peptidoglycan and wall teichoic acid biogenesis pathways. Curr Opin Struct Biol 2018; 53:45–58 [View Article] [PubMed]
     [Google Scholar]
  21. Akhavan BJ, Khanna NR, Amoxicillin VP. Amoxicillin. In StatPearls Treasure Island (FL): StatPearls Publishing; 2023
     [Google Scholar]
  22. Qureshi NN, Morikis D, Schiller NL. Contribution of specific amino acid changes in penicillin binding protein 1 to amoxicillin resistance in clinical Helicobacter pylori isolates. Antimicrob Agents Chemother 2011; 55:101–109 [View Article] [PubMed]
     [Google Scholar]
  23. Gerrits MM, Godoy APO, Kuipers EJ, Ribeiro ML, Stoof J et al. Multiple mutations in or adjacent to the conserved penicillin-binding protein motifs of the penicillin-binding protein 1A confer amoxicillin resistance to Helicobacter pylori. Helicobacter 2006; 11:181–187 [View Article] [PubMed]
     [Google Scholar]
  24. Nagai K, Davies TA, Jacobs MR, Appelbaum PC. Effects of amino acid alterations in penicillin-binding proteins (PBPs) 1a, 2b, and 2x on PBP affinities of penicillin, ampicillin, amoxicillin, cefditoren, cefuroxime, cefprozil, and cefaclor in 18 clinical isolates of penicillin-susceptible, -intermediate, and -resistant pneumococci. Antimicrob Agents Chemother 2002; 46:1273–1280 [View Article] [PubMed]
     [Google Scholar]
  25. Alcock BP, Huynh W, Chalil R, Smith KW, Raphenya AR et al. CARD 2023: expanded curation, support for machine learning, and resistome prediction at the comprehensive antibiotic resistance database. Nucleic Acids Res 2023; 51:D690–D699 [View Article] [PubMed]
     [Google Scholar]
  26. Gerrits MM, Schuijffel D, van Zwet AA, Kuipers EJ, Vandenbroucke-Grauls CMJE et al. Alterations in penicillin-binding protein 1A confer resistance to beta-lactam antibiotics in Helicobacter pylori. Antimicrob Agents Chemother 2002; 46:2229–2233 [View Article] [PubMed]
     [Google Scholar]
  27. Kuo C-J, Ke J-N, Kuo T, Lin C-Y, Hsieh S-Y et al. Multiple amino acid substitutions in penicillin-binding protein-1A confer amoxicillin resistance in refractory Helicobacter pylori infection. J Microbiol Immunol Infect 2023; 56:40–47 [View Article] [PubMed]
     [Google Scholar]
  28. Subsomwong P, Doohan D, Fauzia KA, Akada J, Matsumoto T et al. Next-generation sequencing-based study of Helicobacter pylori isolates from myanmar and their susceptibility to antibiotics. Microorganisms 2022; 10:196 [View Article] [PubMed]
     [Google Scholar]
  29. Francesco VD, Zullo A, Manta R, Satriano A, Fiorini G et al. Culture-based antibiotic susceptibility testing for Helicobacter pylori infection: A systematic review. Ann Gastroenterol 2022; 35:127–134 [View Article] [PubMed]
     [Google Scholar]
  30. Puah SM, Goh KL, Ng HK, Chua KH. Current status of Helicobacter pylori resistance to Clarithromycin and Levofloxacin in Malaysia-findings from a molecular based study. PeerJ 2021; 9:e11518 [View Article] [PubMed]
     [Google Scholar]
  31. Hwong-Ruey Leow A, Chang JV, Goh KL. Searching for an optimal therapy for H pylori eradication: high-dose proton-pump inhibitor dual therapy with amoxicillin vs. standard triple therapy for 14 days. Helicobacter 2020; 25:e12723 [View Article] [PubMed]
     [Google Scholar]
  32. Goh K-L, Lee YY, Leow AH-R, Ali RAR, Ho SH et al. A Malaysian consensus report on the diagnosis and treatment of Helicobacter pylori infection. JGH Open 2023; 7:261–271 [View Article] [PubMed]
     [Google Scholar]
  33. Ng HK, Goh KL, Chuah KH, Thalha AM, Kee BP et al. Sequencing-based detection of 23S rRNA domain V mutations in treatment-naïve Helicobacter pylori patients from Malaysia. J Glob Antimicrob Resist 2020; 23:345–348 [View Article] [PubMed]
     [Google Scholar]
  34. Hall TA. Bioedit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symp Ser 1999; 41:95–98
     [Google Scholar]
  35. Tamura K, Stecher G, Kumar S. MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol 2021; 38:3022–3027 [View Article] [PubMed]
     [Google Scholar]
  36. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987; 4:406–425 [View Article] [PubMed]
     [Google Scholar]
  37. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 1980; 16:111–120 [View Article] [PubMed]
     [Google Scholar]
  38. Letunic I, Bork P. Interactive Tree Of Life (iTOL) v5: an online tool for phylogenetic tree display and annotation. Nucleic Acids Res 2021; 49:W293–W296 [View Article] [PubMed]
     [Google Scholar]
  39. Xu X, He C, Zhu Y. Treatment of refractory Helicobacter pylori infection: a new challenge for clinicians. Front Microbiol 2022; 13:998240 [View Article]
     [Google Scholar]
  40. Mahachai V, Vilaichone R-K, Pittayanon R, Rojborwonwitaya J, Leelakusolvong S et al. Helicobacter pylori management in ASEAN: the Bangkok consensus report. J Gastroenterol Hepatol 2018; 33:37–56 [View Article] [PubMed]
     [Google Scholar]
  41. Tuan VP, Narith D, Tshibangu-Kabamba E, Dung HDQ, Viet PT et al. A next-generation sequencing-based approach to identify genetic determinants of antibiotic resistance in Cambodian Helicobacter pylori clinical isolates. J Clin Med 2019; 8:858 [View Article] [PubMed]
     [Google Scholar]
  42. Zhou Y, Zhong Z, Hu S, Wang J, Deng Y et al. A survey of Helicobacter pylori antibiotic-resistant genotypes and strain lineages by whole-genome sequencing in China. Antimicrob Agents Chemother 2022; 66:e0218821 [View Article] [PubMed]
     [Google Scholar]
  43. Domanovich-Asor T, Motro Y, Khalfin B, Craddock HA, Peretz A et al. Genomic analysis of antimicrobial resistance genotype-to-phenotype agreement in Helicobacter pylori. Microorganisms 2020; 9:2 [View Article] [PubMed]
     [Google Scholar]
  44. Kwon DH, Dore MP, Kim JJ, Kato M, Lee M et al. High-level beta-lactam resistance associated with acquired multidrug resistance in Helicobacter pylori. Antimicrob Agents Chemother 2003; 47:2169–2178 [View Article] [PubMed]
     [Google Scholar]
  45. Tomb JF, White O, Kerlavage AR, Clayton RA, Sutton GG et al. The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature 1997; 388:539–547 [View Article] [PubMed]
     [Google Scholar]
  46. Alm RA, Ling LS, Moir DT, King BL, Brown ED et al. Genomic-sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori. Nature 1999; 397:176–180 [View Article] [PubMed]
     [Google Scholar]
  47. Matteo MJ, Granados G, Olmos M, Wonaga A, Catalano M. Helicobacter pylori amoxicillin heteroresistance due to point mutations in PBP-1A in isogenic isolates. J Antimicrob Chemother 2008; 61:474–477 [View Article] [PubMed]
     [Google Scholar]
  48. Zerbetto De Palma G, Mendiondo N, Wonaga A, Viola L, Ibarra D et al. Occurrence of mutations in the antimicrobial target genes related to levofloxacin, clarithromycin, and amoxicillin resistance in Helicobacter pylori isolates from Buenos Aires City. Microb Drug Resist 2017; 23:351–358 [View Article] [PubMed]
     [Google Scholar]
  49. Saranathan R, Levi MH, Wattam AR, Malek A, Asare E et al. Helicobacter pylori infections in the Bronx, New York: surveying antibiotic susceptibility and strain lineage by whole-genome sequencing. J Clin Microbiol 2020; 58:e01591-19 [View Article] [PubMed]
     [Google Scholar]
  50. Tshibangu-Kabamba E, Ngoma-Kisoko P de J, Tuan VP, Matsumoto T, Akada J et al. Next-generation sequencing of the whole bacterial genome for tracking molecular insight into the broad-spectrum antimicrobial resistance of Helicobacter pylori clinical isolates from the democratic Republic of Congo. Microorganisms 2020; 8:887 [View Article] [PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001832
Loading
Genetic variations of penicillin-binding protein 1A: insights into the current status of amoxicillin-based regimens for Helicobacter pylori eradication in Malaysia
J. Med. Microbiol. 73, 001832 (2024); https://doi.org/10.1099/jmm.0.001832
/content/journal/jmm/10.1099/jmm.0.001832
/content/journal/jmm/10.1099/jmm.0.001832
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error