1887

Abstract

The molecular mechanism of resistance to tetracycline involves mutations in the primary binding site of the ribosome. A resistance or reduced susceptibility to tetracycline could be the result of single, double or triple mutations in the 16S gene of . We investigated if the genotype was correlated to tetracycline resistance as determined phenotypically for 96 . isolates in the gastroesophageal mucosa of Venezuelan individual hosts. E-test for antimicrobial susceptibility test and real-time PCR for the detection of 16S gene mutations were performed in 96 . isolates (48 obtained from antrum, and 48 from oesophagus) from eight dyspeptic patients. In the gastric mucosa, 38 isolates were identified sensitive and 10 resistant to tetracycline by E-test, whereas 44 sensitive and 4 resistant isolates were found in the oesophagus. Real-time PCR detection of the 16S gene exhibited mutants with a single base-pair substitution (AGA GA) in six antrum isolates and seven oesophagus isolates, whereas only three harboured a low level of tetracycline resistance . Our results indicate that real-time PCR detection of 16S is a reliable method to classify among tetracycline-resistant genotypes and useful in patients who have experienced a first-line treatment failure with triple therapy.

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2019-09-01
2019-10-17
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References

  1. Peña J, Rojas H, Reyes N, Fernández-Delgado M, García-Amado M-A et al. Multiple cag genotypes of Helicobacter pylori isolates colonize the oesophagus in individual hosts in a Venezuelan population. J Med Microbiol 2017;66:226–235 [CrossRef]
    [Google Scholar]
  2. Hunt RH, Xiao SD, Megraud F, Leon-Barua R, Bazzoli F et al. Helicobacter pylori in developing countries. World Gastroenterology Organisation global guidelines of H. pylori. WGO 2010
    [Google Scholar]
  3. Seck A, Burucoa C, Dia D, Mbengue M, Onambele M et al. Primary antibiotic resistance and associated mechanisms in Helicobacter pylori isolates from Senegalese patients. Ann Clin Microbiol Antimicrob 2013;12:3–7 [CrossRef]
    [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 [CrossRef]
    [Google Scholar]
  5. Gerrits MM, de Zoete MR, Arents NLA, Kuipers EJ, Kusters JG. 16S rRNA mutation-mediated tetracycline resistance in Helicobacter pylori. Antimicrob Agents Chemother 2002;46:2996–3000 [CrossRef]
    [Google Scholar]
  6. Trieber CA, Taylor DE. Mutations in the 16S rRNA genes of Helicobacter pylori mediate resistance to tetracycline. J Bacteriol 2002;184:2131–2140 [CrossRef]
    [Google Scholar]
  7. Glocker E, Berning M, Gerrits MM, Kusters JG, Kist M. Real-Time PCR screening for 16S rRNA mutations associated with resistance to tetracycline in Helicobacter pylori. Antimicrob Agents Chemother 2005;49:3166–3170 [CrossRef]
    [Google Scholar]
  8. Contreras M, Abrante L, Salazar V, Reyes N, García-Amado MA et al. Heterogeneity of cag genotypes of Helicobacter pylori in the esophageal mucosa of dyspeptic patients and its relation to histopathological outcomes. Int J Infect Dis 2014;26:91–95 [CrossRef]
    [Google Scholar]
  9. Domínguez-Bello MG, Michelangeli F, Romero R, Beker B, Lara D et al. Modification of Christensen urease test as an inexpensive tool for detection of Helicobacter pylori. Diagn Microbiol Infect Dis 1997;28:149–152 [CrossRef]
    [Google Scholar]
  10. Germani Y, Dauga C, Duval P, Huerre M, Levy M et al. Strategy for the detection of Helicobacter species by amplification of 16S rRNA genes and identification of H. felis in a human gastric biopsy. Res Microbiol 1997;148:315–326 [CrossRef]
    [Google Scholar]
  11. Kansau I, Raymond J, Bingen E, Courcoux P, Kalach N et al. Genotyping of Helicobacter pylori isolates by sequencing of PCR products and comparison with the RAPD technique. Res Microbiol 1996;147:661–669 [CrossRef]
    [Google Scholar]
  12. Peek RM, Miller GG, Tham KT, Pérez-Pérez GI, Cover TL et al. Detection of Helicobacter pylori gene expression in human gastric mucosa. J Clin Microbiol 1995;33:28–32
    [Google Scholar]
  13. Dadashzadeh K, Milani M, Rahmati M, Akbarzadeh A. Real-Time PCR detection of 16S rRNA novel mutations associated with Helicobacter pylori tetracycline resistance in Iran. Asian Pac J Cancer Prev 2014;15:8883–8886 [CrossRef]
    [Google Scholar]
  14. Seriki AT, Smith SI, Adeleye AI, Fowora MA. Molecular analysis of low-level tetracycline resistance in clinical isolates of Helicobacter pylori among dyspeptic patients in South West Nigeria. J Glob Antimicrob Resist 2018;13:143–145 [CrossRef]
    [Google Scholar]
  15. Bachir M, Allem R, Benejat L, Tifrit A, Medjekane M et al. Molecular detection of mutations involved in Helicobacter pylori antibiotic resistance in Algeria. J Antimicrob Chemother 2018;73:2034–2038 [CrossRef]
    [Google Scholar]
  16. Lawson AJ, Elviss NC, Owen RJ. Real-Time PCR detection and frequency of 16S rDNA mutations associated with resistance and reduced susceptibility to tetracycline in Helicobacter pylori from England and Wales. J Antimicrob Chemother 2005;56:282–286 [CrossRef]
    [Google Scholar]
  17. López-Gasca M, García-Amado M-A, Michelangeli F, Contreras M, Peña J. Point mutations at gyrA and gyrB genes of Levofloxacin-Resistant Helicobacter pylori isolates in the esophageal mucosa from a Venezuelan population. Am J Trop Med Hyg 2018;98:1051–1055 [CrossRef]
    [Google Scholar]
  18. Cuchí Burgos E, Forné Bardera M, Quintana Riera S, Lite Lite J, Garau Alemany J. [Evolution of the sensitivity of 235 strains of Helicobacter pylori from 1995 to 1998 and impact of antibiotic treatment]. Enferm Infecc Microbiol Clin 2002;20:157–160
    [Google Scholar]
  19. Kim JJ, Reddy R, Lee M, Kim JG, El-Zaatari FA et al. Analysis of metronidazole, clarithromycin and tetracycline resistance of Helicobacter pylori isolates from Korea. J Antimicrob Chemother 2001;47:459–461 [CrossRef]
    [Google Scholar]
  20. Parsons HK, Carter MJ, Sanders DS, Winstanley T, Lobo AJ. Helicobacter pylori antimicrobial resistance in the United Kingdom: the effect of age, sex and socio-economic status. Aliment Pharmacol Ther 2001;15:1473–1478 [CrossRef]
    [Google Scholar]
  21. Kwon DH, Kim JJ, Lee M, Yamaoka Y, Kato M et al. Isolation and characterization of tetracycline-resistant clinical isolates of Helicobacter pylori. Antimicrob Agents Chemother 2000;44:3203–3205 [CrossRef]
    [Google Scholar]
  22. Realdi G, Dore MP, Piana A, Atzei A, Carta M et al. Pretreatment antibiotic resistance in Helicobacter pylori infection: results of three randomized controlled studies. Helicobacter 1999;4:106–112 [CrossRef]
    [Google Scholar]
  23. Boyanova L, Mitov I. Geographic map and evolution of primary Helicobacter pylori resistance to antibacterial agents. Expert Rev Anti Infect Ther 2010;8:59–70 [CrossRef]
    [Google Scholar]
  24. Wu JY, Kim JJ, Reddy R, Wang WM, Graham DY et al. Tetracycline-Resistant clinical Helicobacter pylori isolates with and without mutations in 16S rRNA-encoding genes. Antimicrob Agents Chemother 2005;49:578–583 [CrossRef]
    [Google Scholar]
  25. Ducournau A, Bénéjat L, Sifré E, Bessède E, Lehours P et al. Helicobacter pylori resistance to antibiotics in 2014 in France detected by phenotypic and genotypic methods. Clin Microbiol Infect 2016;22:715–718 [CrossRef]
    [Google Scholar]
  26. Camargo MC, García A, Riquelme A, Otero W, Camargo CA et al. The problem of Helicobacter pylori resistance to antibiotics: a systematic review in Latin America. Am J Gastroenterol 2014;109:485–495 [CrossRef]
    [Google Scholar]
  27. Mendonça S, Ecclissato C, Sartori MS, Godoy APO, Guerzoni RA et al. Prevalence of Helicobacter pylori resistance to metronidazole, clarithromycin, amoxicillin, tetracycline, and furazolidone in Brazil. Helicobacter 2000;5:79–83 [CrossRef]
    [Google Scholar]
  28. Vallejos C, Garrido L, Cáceres D, Madrid AM, Defilippi C et al. [Prevalence of metronidazole, clarithromycin and tetracycline resistance in Helicobacter pylori isolated from Chilean patients]. Rev Med Chil 2007;135:287–293
    [Google Scholar]
  29. Urrestarazu M, Serrano N, Piñero R, Cavazza M. Susceptibilidad de Helicobacter pylori a los antimicrobianos/Antimicrobial susceptibility of Helicobacter pylori. Rev Soc Venez Microbiol 2003;23:14–15
    [Google Scholar]
  30. Chu YT, Wang YH, Wu JJ, Lei HY. Invasion and multiplication of Helicobacter pylori in gastric epithelial cells and implications for antibiotic resistance. Infect Immun 2010;78:4157–4165 [CrossRef]
    [Google Scholar]
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