Zanco Journal of Medical Sciences
Skip Navigation Links
Available Issues
Editorial Board
Information for Authors
Review Process
Links and Contacts
  Zanko J Med Sci:  Apr. 2017; 21 (1): 1653-1658

Molecular detection of Helicobacter species in the bile and histopathological evaluation of gall bladders of patients with calculus cholycystitis in Duhok province, Kurdistan region of Iraq

Amir Hani. Raziq*, Alaa Hani. Raziq**, Sardar Hassan Arif***

* Department of Engineering and biotechnology, College of Sciences, Duhok University, Duhok, Iraq.

** Department of Pathology, College of Medicine, Duhok University, Duhok, Iraq.

*** Department of Surgery, College of Medicine, Duhok University, Duhok, Iraq.


Background and objective: The presence of bile-resistant Helicobacter spp. in bile and gallbladder tissues has been proposed as a cause of gallbladder disease. This study aimed to assess the histopathological events in calculus cholecystitis and investigate the presence of the DNA of Helicobacter spp. in the bile.   

Methods: Forty patients who underwent cholecystectomy for calculus cholecystitis were evaluated for the presence of Helicobacter spp. by polymerase chain reaction methods and gallbladder tissue was evaluated histopathologically. Gall bladders were fixed in formalin and subjected to histopathological investigation while bile sample was used for the extraction of DNA.

Results: The histopathological findings revealed that twenty-seven (67.5%) samples showed mild chronic inflammation, one (2.5 %) sample with acute inflammation, four (10 %) samples with acute on chronic inflammation, eight (20 %) samples have normal histological findings. The molecular investigation revealed that twenty one (52 %) samples out of the forty showed the presence of genomic DNA after being subjected to DNA extraction and that fourteen (66.7 %) samples out of the twenty one showed amplification bands of 300 bp after PCR amplification indicating the presence of Helicobacter spp. within the samples. But no sample revealed the presence of H. pylori DNA when PCR was applied.   

Conclusion: Helicobacter spp. are considered as a determining factor for the development of gall bladder inflammation. 

Keywords: Helicobacter spp.; Gallbladder; Bile; PCR; 16 S rRNA; HPU1; HPU2.


1. Fox JG. The non-H. pylorihelicobacters: their expanding role in gastrointestinal and systemic diseases. Gut 2002; 50:273–83.

2. Kusters JG, VanVliet AHM, Kuipers EJ. Pathogenesis of Helicobacter pylori Infection. Clin Microbiol Reviews 2006; 19(3):449–90.

3. García A, Zeng Y, Muthupalani S, Ge Z, Potter A, Mobley MW, et al. Helicobacter hepaticus–induced liver tumor promotion is associated with increased serum bile acid and a persistent microbial-induced immune response. Cancer Res 2011; 71(7):2529–40.

4. Nillson HO, Taneera J, Castedal M, Glatz E, Olsson R, Wadstrom T. Identification of Helicobacter pylori and other Helicobacter species by PCR, hybridization and partial DNA sequencing in human liver samples from patients with primary sclerosing cholangitis or primary biliary cirrhosis. J Clin Microbiol 2000; 38:1072–6.

5. Swidsinski A, Ludwig W, Pahlig H, Priem F. Molecular genetic evidence of bacterial colonization of cholesterol gallstones. Gastroenterology 1995; 108(3):860–4.

6. Nilsson B, Friman S, Thune A, Jivegard L, Svanvik J. Inflammation reduces mucosal secretion of hydrogen ions and impairs concentrating function and luminal acidification in feline gallbladder. Scand J Gastroenterol 1995; 30:1021–6.

7. Cetta F, Figura N, Montalto G, Cetta D, Baldi C, Amoroso M, et al. Helicobacter pylori in the bile could play a facilitating role in the pathogenesis of the some types of gallstones. Gastroenterology 2000; 118:A7.

8. Pandey M. Helicobacter species are associated with possible increase in risk of biliary lithiasis and benign biliary diseases. World J Surg Oncol 2007; 5:94.

9. Rosai J. Rosai and Ackerman’s surgical pathology. New York: Elsevier Corporation; 2011.

10. Bancroft JD, Steven A. Theory and practice of histopathological techniques. Edinburgh Churchill Livingstones; 1999.

11. Silva CP, Pereira-Lima JC, Oliveira AG, Guerra JB, Marques DL, Sarmanho L, et al. Association of the presence of Helicobacter in gallbladder tissue with cholelithiasis and cholecystitis. J Clin Microbiol 2003; 41:5615-8.

12. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: A laboratory manual. New York: Cold spring harbor laboratory press; 1989.

13. Ward JM, Anver MR, Haines DC, Benveniste RE. Chronic active hepatitis in mice caused by Helicobacter hepaticus. Am J Pathol 1994; 145:959–68.

14. Ghazal A, El Sabbagh N, El Riwini M. Presence of Helicobacter spp. DNA in the gallbladder of Egyptian patients with gallstone diseases. East Mediterr Health J 2011; 17(12):925–9.

15. Lee JW, Lee DH, Lee JI, Jeong S, Kwon KS, Kim HG, et al. Identification of Helicobacter pylori in gallstone, bile, and other hepatobiliary tissues of patients with cholecystitis. Gut and Liver 2010; 4:60–7.

16. Tiwari SK, Khan AA, Ibrahim M, Habeeb MA, Habibullah CM. Helicobacter pylori and other Helicobacter species DNA in human bile samples from patients with various hepato-biliary diseases. World J Gastroenterol 2006; 12:2181–6.

17. Fox JG, Dewhirst FE, Shen Z. Hepatic Helicobacter species identified in the bile and gallbladder tissue from Chileans with chronic cholecystitis. Gastroenterology 1998; 114:755–63.

18. Fallone CA, Tran S, Semret M, Discepola F,   Behr M, Barkun AN. Helicobacter DNA in bile: correlation with hepato-biliary diseases. Aliment Pharmacol Ther 2003; 17:453–8.

19. Chen DF, Hu L, Yi P, Liu WW, Fang DC, Cao H. H. pylori exist in the gallbladder mucosa of patients with chronic cholecystitis. World J Gastroenterol 2007; 13:1608–11. 

20. Rudi J, Rudy A, Maiwald M, Stremmel W. Helicobacter sp. are not detectable in bile from German patients with biliary disease. Gastroenterology 1999; 116:1016–7.

21. Nilsson B, Friman S, Thune A, Jivegard L, Svanvik J. Inflammation reduces mucosal secretion of hydrogen ions and impairs concentrating function and luminal acidification in feline gallbladder. Scand J Gastroenterol 1995; 30:1021–6.

22. Erden E, Kiyan M. Evaluation of the presence of Helicobacter species in the biliary system of Turkish patients with cholelithiasis. Turk J Gastroenterol 2010; 21:421-427.

23. Pellicano R, Menard A, Rizzetto M, Megraud F. Helicobacter species and liver diseases: association or causation? Lancet Infect Dis 2008; 8(4):254–60.

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.