Concurrent detection of cagA, vacA, sodB and hsp60 virulence genes and their relationship with clinical outcomes of disease in Helicobacter pylori isolated strains of southwest of Iran
Abstract
Background and Objectives: Helicobacter pylori is a Gram-negative spiral-shaped bacterium that contaminates more than half of the world's inhabitants, and infection with this bacterium is associated with some gastric disorders. Also, 5% to 10% of H. pylori genes are specific to this bacterium and many bacterial virulence factors fall into this group. The cagA, vacA, sodB and hsp60 are among important virulence factors of H. pylori.
Materials and Methods: A gastric biopsy specimen was taken from 341 gastric patients and cultivated on a Colombia agar plate, containing various antibiotics, such as vancomycin, amphotericin B, and trimethoprim & polymyxin B, and incubated for 3 to 10 days under microaerophilic conditions at 37°C. PCR was used to detect the ureC, cagA, vacA, sodB and hsp60 genes.
Results: In this study, 131 isolates were identified as H. pylori. The prevalence of cagA, vacA, sodB and hsp60 were 74%, 100%, 92.4% and 96.2%, respectively. The correlation between the clinical forms of the disease and the virulence genes were analyzed by statistical tests and no significant correlation was found.
Conclusion: The obtained results are similar to some studies conducted in different parts of the world and is different in other cases. This discrepancy is due to the difference in the type of gastric disorders, sample size and methodology.
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2. Bridge DR, Merrell DS. Polymorphism in the Helicobacter pylori CagA and VacA toxins and disease. Gut Microbes 2013;4:101-117.
3. Salama NR, Hartung ML, Müller A. Life in the human stomach: persistence strategies of the bacterial pathogen Helicobacter pylori. Nat Rev Microbiol 2013;11:385-399.
4. Rasheed F, Campbell BJ, Alfizah H, Varro A, Zahra R, Yamaoka Y, et al. Analysis of clinical isolates of Helicobacter pylori in Pakistan reveals high degrees of pathogenicity and high frequencies of antibiotic resistance. Helicobacter 2014;19:387-399.
5. Santos J, Ladeira M, Pedrazzoli Jr J, Ribeiro M. Relationship of IL-1 and TNF-α polymorphisms with Helicobacter pylori in gastric diseases in a Brazilian population. Braz J Med Biol Res 2012;45:811-817.
6. Vaziri F, Peerayeh SN, Alebouyeh M, Mirzaei T, Yamaoka Y, Molaei M, et al. Diversity of Helicobacter pylori genotypes in Iranian patients with different gastroduodenal disorders. World J Gastroenterol 2013;19:5685-5692.
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8. Menoni SM, Bonon SH, Zeitune JM, Costa SC. PCR-based detection and genotyping of Helicobacter pylori in endoscopic biopsy samples from Brazilian patients. Gastroenterol Res Pract 2013; 2013:951034.
9. Owen RJ, Xerry J, Gotada T, Naylor G, Tompkins D. Analysis of geospecific markers for Helicobacter pylori variants in patients from Japan and Nigeria by triple-locus nucleotide sequence typing. Microbiology 2004;150:151-161.
10. Tegtmeyer N, Wessler S, Backert S. Role of the cag-pathogenicity island encoded type IV secretion system in Helicobacter pylori pathogenesis. FEBS J 2011;278:1190-1202.
11. Yamaoka Y, Graham DY. Helicobacter pylori virulence and cancer pathogenesis. Future Oncol 2014;10:1487-1500.
12. Polk DB, Peek Jr RM. Helicobacter pylori: gastric cancer and beyond. Nat Rev Cancer 2010;10:403-414.
13. Ayala G, Escobedo-Hinojosa WI, de la Cruz-Herrera CF, Romero I. Exploring alternative treatments for Helicobacter pylori infection. World J Gastroenterol 2014;20:1450-1469.
14. Saribasak H, Salih BA, Yamaoka Y, Sander E. Analysis of Helicobacter pylori genotypes and correlation with clinical outcome in Turkey. J Clin Microbiol 2004;42:1648-1651.
15. Yong X, Tang B, Li B-S, Xie R, Hu C-J, Luo G, et al. Helicobacter pylori virulence factor CagA promotes tumorigenesis of gastric cancer via multiple signaling pathways. Cell Commun Signal 2015;13:30.
16. Yamaoka Y. Mechanisms of disease: Helicobacter pylori virulence factors. Nat Rev Gastroenterol Hepatol 2010;7:629-641.
17. Maleki P, Latifi-Navid S, Zahri S. Allelic variation of Helicobacter pylori babA and cagA genes and their association with clinical consequences. Govaresh 2013;17:203-212.
18. Latifi-Navid S, Mohammadi S, Maleki P, Zahri S, Yazdanbod A, Siavoshi F, et al. Helicobacter pylori vacA d1/-i1 genotypes and geographic differentiation between high and low incidence areas of gastric cancer in Iran. Arch Iran Med 2013;16:330-337.
19. Chattopadhyay S, Patra R, Ramamurthy T, Chowdhury A, Santra A, Dhal GK, et al. Multiplex PCR assay for rapid detection and genotyping of Helicobacter pylori directly from biopsy specimens. J Clin Microbiol 2004;42: 2821-2824.
20. Rajabnia R, Salehi M. Outbreak of Caga and Icea in H. Pylori strains isolated from patients with gastro duodenal diseases in Babol city. Iran J Public Health 2014;43:189-196.
21. Blaser MJ, Atherton JC. Helicobacter pylori persistence: biology and disease. J Clin Invest 2004;113:321-333.
22. Schraw W, Li Y, McClain MS, van der Goot FG, Cover TL. Association of Helicobacter pylori vacuolating toxin (VacA) with lipid rafts. J Biol Chem 2002;277:34642-34650.
23. Palframan SL, Kwok T, Gabriel K. Vacuolating cytotoxin A (VacA), a key toxin for Helicobacter pylori pathogenesis. Front Cell Infect Microbiol 2012; 2: 92.
24. Raju D, Hussey S, Ang M, Terebiznik MR, Sibony M, Galindo–Mata E, et al. Vacuolating cytotoxin and variants in Atg16L1 that disrupt autophagy promote Helicobacter pylori infection in humans. Gastroenterology 2012;142:1160-1171.
25. A EV, TI Ca, O NlP, H JsS. Molecular characterization and susceptibility testing of Helicobacter pylori strains isolated in western Argentina. Int J Infect Dis 2010;14:e85-e92.
26. Takeshima E, Tomimori K, Takamatsu R, Ishikawa C, Kinjo F, Hirayama T, et al. Helicobacter pylori VacA activates NF-kappaB in T cells via the classical but not alternative pathway. Helicobacter 2009;14:271-279.
27. Ryberg A, Borch K, Sun Y-Q, Monstein H-J. Concurrent genotyping of Helicobacter pylori virulence genes and human cytokine SNP sites using whole genome amplified DNA derived from minute amounts of gastric biopsy specimen DNA. BMC Microbiol 2008;8:175.
28. Slonczewski JL, McGee DJ, Phillips J, Kirkpatrick C, Mobley HL. pH-dependent protein profiles of Helicobacter pylori analyzed by two-dimensional gels. Helicobacter 2000;5:240-247.
29. Jungblut P, Bumann D, Haas G, Zimny-Arndt U, Holland P, Lamer S, et al. Comparative proteome analysis of Helicobacter pylori. Mol Microbiol 2000;36:710-725.
30. Mendoza JA, Weinberger KK, Swan MJ. The Hsp60 protein of Helicobacter pylori displays chaperone activity under acidic conditions. Biochem Biophys Rep 2017;9:95-99.
31. Gobert AP, Bambou J-C, Werts C, Balloy V, Chignard M, Moran AP, et al. Helicobacter pylori heat shock protein 60 mediates interleukin-6 production by macrophages via a toll-like receptor (TLR)-2-, TLR-4-, and myeloid differentiation factor 88-independent mechanism. J Biol Chem 2004;279:245-250.
32. Forbes B. Weissfeld AS. Bailey&Scotts Dia microbiology. USA, Elsevier; 2007.
33. Zhao L-J, Huang Y-Q, Chen B-P, Mo X-Q, Huang Z-S, Huang X-F, et al. Helicobacter pylori isolates from ethnic minority patients in Guangxi: Resistance rates, mechanisms, and genotype. World J Gastroenterol 2014;20:4761-4770.
34. Tanih NF, Ndip RN. Molecular detection of antibiotic resistance in South African isolates of Helicobacter pylori. Gastroenterol Res Pract 2013; 2013: 259457.
35. Farshad S, Japoni A, Alborzi A, Hosseini M. Restriction fragment length polymorphism of virulence genes cagA, vacA and ureAB of Helicobacter pylori strains isolated from Iranian patients with gastric ulcer and nonulcer disease. Saudi Med J 2007;28:529-534.
36. Shiota S, Cruz M, Abreu JAJ, Mitsui T, Terao H, Disla M, et al. Virulence genes of Helicobacter pylori in the Dominican Republic. J Med Microbiol 2014;63:1189-1196.
2. Bridge DR, Merrell DS. Polymorphism in the Helicobacter pylori CagA and VacA toxins and disease. Gut Microbes 2013;4:101-117.
3. Salama NR, Hartung ML, Müller A. Life in the human stomach: persistence strategies of the bacterial pathogen Helicobacter pylori. Nat Rev Microbiol 2013;11:385-399.
4. Rasheed F, Campbell BJ, Alfizah H, Varro A, Zahra R, Yamaoka Y, et al. Analysis of clinical isolates of Helicobacter pylori in Pakistan reveals high degrees of pathogenicity and high frequencies of antibiotic resistance. Helicobacter 2014;19:387-399.
5. Santos J, Ladeira M, Pedrazzoli Jr J, Ribeiro M. Relationship of IL-1 and TNF-α polymorphisms with Helicobacter pylori in gastric diseases in a Brazilian population. Braz J Med Biol Res 2012;45:811-817.
6. Vaziri F, Peerayeh SN, Alebouyeh M, Mirzaei T, Yamaoka Y, Molaei M, et al. Diversity of Helicobacter pylori genotypes in Iranian patients with different gastroduodenal disorders. World J Gastroenterol 2013;19:5685-5692.
7. Ferreira RM, Machado JC, Letley D, Atherton JC, Pardo ML, Gonzalez CA, et al. A novel method for genotyping Helicobacter pylori vacA intermediate region directly in gastric biopsy specimens. J Clin Microbiol 2012; 50: 3983-3989.
8. Menoni SM, Bonon SH, Zeitune JM, Costa SC. PCR-based detection and genotyping of Helicobacter pylori in endoscopic biopsy samples from Brazilian patients. Gastroenterol Res Pract 2013; 2013:951034.
9. Owen RJ, Xerry J, Gotada T, Naylor G, Tompkins D. Analysis of geospecific markers for Helicobacter pylori variants in patients from Japan and Nigeria by triple-locus nucleotide sequence typing. Microbiology 2004;150:151-161.
10. Tegtmeyer N, Wessler S, Backert S. Role of the cag-pathogenicity island encoded type IV secretion system in Helicobacter pylori pathogenesis. FEBS J 2011;278:1190-1202.
11. Yamaoka Y, Graham DY. Helicobacter pylori virulence and cancer pathogenesis. Future Oncol 2014;10:1487-1500.
12. Polk DB, Peek Jr RM. Helicobacter pylori: gastric cancer and beyond. Nat Rev Cancer 2010;10:403-414.
13. Ayala G, Escobedo-Hinojosa WI, de la Cruz-Herrera CF, Romero I. Exploring alternative treatments for Helicobacter pylori infection. World J Gastroenterol 2014;20:1450-1469.
14. Saribasak H, Salih BA, Yamaoka Y, Sander E. Analysis of Helicobacter pylori genotypes and correlation with clinical outcome in Turkey. J Clin Microbiol 2004;42:1648-1651.
15. Yong X, Tang B, Li B-S, Xie R, Hu C-J, Luo G, et al. Helicobacter pylori virulence factor CagA promotes tumorigenesis of gastric cancer via multiple signaling pathways. Cell Commun Signal 2015;13:30.
16. Yamaoka Y. Mechanisms of disease: Helicobacter pylori virulence factors. Nat Rev Gastroenterol Hepatol 2010;7:629-641.
17. Maleki P, Latifi-Navid S, Zahri S. Allelic variation of Helicobacter pylori babA and cagA genes and their association with clinical consequences. Govaresh 2013;17:203-212.
18. Latifi-Navid S, Mohammadi S, Maleki P, Zahri S, Yazdanbod A, Siavoshi F, et al. Helicobacter pylori vacA d1/-i1 genotypes and geographic differentiation between high and low incidence areas of gastric cancer in Iran. Arch Iran Med 2013;16:330-337.
19. Chattopadhyay S, Patra R, Ramamurthy T, Chowdhury A, Santra A, Dhal GK, et al. Multiplex PCR assay for rapid detection and genotyping of Helicobacter pylori directly from biopsy specimens. J Clin Microbiol 2004;42: 2821-2824.
20. Rajabnia R, Salehi M. Outbreak of Caga and Icea in H. Pylori strains isolated from patients with gastro duodenal diseases in Babol city. Iran J Public Health 2014;43:189-196.
21. Blaser MJ, Atherton JC. Helicobacter pylori persistence: biology and disease. J Clin Invest 2004;113:321-333.
22. Schraw W, Li Y, McClain MS, van der Goot FG, Cover TL. Association of Helicobacter pylori vacuolating toxin (VacA) with lipid rafts. J Biol Chem 2002;277:34642-34650.
23. Palframan SL, Kwok T, Gabriel K. Vacuolating cytotoxin A (VacA), a key toxin for Helicobacter pylori pathogenesis. Front Cell Infect Microbiol 2012; 2: 92.
24. Raju D, Hussey S, Ang M, Terebiznik MR, Sibony M, Galindo–Mata E, et al. Vacuolating cytotoxin and variants in Atg16L1 that disrupt autophagy promote Helicobacter pylori infection in humans. Gastroenterology 2012;142:1160-1171.
25. A EV, TI Ca, O NlP, H JsS. Molecular characterization and susceptibility testing of Helicobacter pylori strains isolated in western Argentina. Int J Infect Dis 2010;14:e85-e92.
26. Takeshima E, Tomimori K, Takamatsu R, Ishikawa C, Kinjo F, Hirayama T, et al. Helicobacter pylori VacA activates NF-kappaB in T cells via the classical but not alternative pathway. Helicobacter 2009;14:271-279.
27. Ryberg A, Borch K, Sun Y-Q, Monstein H-J. Concurrent genotyping of Helicobacter pylori virulence genes and human cytokine SNP sites using whole genome amplified DNA derived from minute amounts of gastric biopsy specimen DNA. BMC Microbiol 2008;8:175.
28. Slonczewski JL, McGee DJ, Phillips J, Kirkpatrick C, Mobley HL. pH-dependent protein profiles of Helicobacter pylori analyzed by two-dimensional gels. Helicobacter 2000;5:240-247.
29. Jungblut P, Bumann D, Haas G, Zimny-Arndt U, Holland P, Lamer S, et al. Comparative proteome analysis of Helicobacter pylori. Mol Microbiol 2000;36:710-725.
30. Mendoza JA, Weinberger KK, Swan MJ. The Hsp60 protein of Helicobacter pylori displays chaperone activity under acidic conditions. Biochem Biophys Rep 2017;9:95-99.
31. Gobert AP, Bambou J-C, Werts C, Balloy V, Chignard M, Moran AP, et al. Helicobacter pylori heat shock protein 60 mediates interleukin-6 production by macrophages via a toll-like receptor (TLR)-2-, TLR-4-, and myeloid differentiation factor 88-independent mechanism. J Biol Chem 2004;279:245-250.
32. Forbes B. Weissfeld AS. Bailey&Scotts Dia microbiology. USA, Elsevier; 2007.
33. Zhao L-J, Huang Y-Q, Chen B-P, Mo X-Q, Huang Z-S, Huang X-F, et al. Helicobacter pylori isolates from ethnic minority patients in Guangxi: Resistance rates, mechanisms, and genotype. World J Gastroenterol 2014;20:4761-4770.
34. Tanih NF, Ndip RN. Molecular detection of antibiotic resistance in South African isolates of Helicobacter pylori. Gastroenterol Res Pract 2013; 2013: 259457.
35. Farshad S, Japoni A, Alborzi A, Hosseini M. Restriction fragment length polymorphism of virulence genes cagA, vacA and ureAB of Helicobacter pylori strains isolated from Iranian patients with gastric ulcer and nonulcer disease. Saudi Med J 2007;28:529-534.
36. Shiota S, Cruz M, Abreu JAJ, Mitsui T, Terao H, Disla M, et al. Virulence genes of Helicobacter pylori in the Dominican Republic. J Med Microbiol 2014;63:1189-1196.
| Files | ||
| Issue | Vol 11 No 3 (2019) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v11i3.1315 | |
| Keywords | ||
| Helicobacter pylori Multiplex polymerase chain reaction Virulence genes Chronic gastritis Gastric disorders peptic ulcers | ||
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How to Cite
1.
Amin M, Shayesteh AA, Serajian A. Concurrent detection of cagA, vacA, sodB and hsp60 virulence genes and their relationship with clinical outcomes of disease in Helicobacter pylori isolated strains of southwest of Iran. Iran J Microbiol. 2019;11(3):198-205.
