The Prevalence of Helicobacter pylori babA2, iceA1 and iceA2 Genes and Their Association with Clinical Outcomes in Patients with Chronic Gastritis, Ulcerative Diseases and Non-Ulcer Dyspepsia in South East of Iran


avatar Hamid Abdollahi 1 , avatar Mostafa shokoohi 2 , avatar Mohammad Savari 1 , *

Department of Microbiology, Virology & Immunology, School of Medicine, Kerman University of Medical Sciences, Kerman, IR Iran, Kerman, IR Iran
Kerman Physiology Research Center (KPRC), Kerman University of Medical Sciences, Kerman, IR Iran

how to cite: Abdollahi H, shokoohi M, Savari M. The Prevalence of Helicobacter pylori babA2, iceA1 and iceA2 Genes and Their Association with Clinical Outcomes in Patients with Chronic Gastritis, Ulcerative Diseases and Non-Ulcer Dyspepsia in South East of Iran. Jundishapur J Microbiol. 2013;6(4):e4739.



Helicobacter pylori virulence factors are important in development of the clinical outcomes. The initial stage of colonization is binding of H. pylori to gastric epithelial cells through the babA protein. Heterogeneity among H. pylori strains in presence and expressing the babA gene may be a factor in the variation of clinical outcomes. Likewise, another recently H. pylori described virulence factor; iceA has been shown to be a marker for ulcerative diseases.


We investigated the presence of babA2, iceA1 and iceA2H. pylori virulence factors in patients with clinical outcomes in the southeast of Iran.

Patients and Methods:

In this study, 63 positive culture samples out of 191 biopsies examined to determine of babA2, iceA1 and iceA2 genes by PCR. DNA extracted from 63 Helicobacter positive specimens including 46 chronic active gastritis, 6 ulcerative diseases and non-ulcer dyspepsia 11.


The frequency of the babA2, iceA1 and iceA2 genes in the total isolates were 34 (54%), 14 (22.2%) and 34 (54%), respectively. The association of these virulence factors based on sex and age groups were not statistically significant (P > 0.05). There was a borderline significant association between iceA1 and the clinical outcomes (P = 0.094).

2. Objectives

The aim of this present study was to determine the frequency of babA2, iceA1 & iceA2 virulence factors among our local H. pylori isolates and their association with the gastric outcomes, including: chronic gastritis, ulcerative disease such as peptic ulcer disease, duodenal ulcer disease and non- ulcer dyspepsia (NUD).

3. Patients and Methods

Sixty-three H. pylori isolates were obtained from 191 patients' biopsy samples referred to the endoscopy unit of Afzalipour hospital in Kerman, (southeast of Iran) during 2009. The biopsy samples were cultivated in Brucella agar medium (Merck, Germany), supplemented with 10% defibrinated sheep blood and three antibiotics (vancomycin 10mg/L, amphotericin B 10mg/L and trimetoprim 5mg/L). The inoculated plates were incubated at 37°C under microaerophilic atmosphere provided by anerocult C (Merck, Germany) for 3-5 days. The isolates were recognized as H. pylori by urease, catalase, oxidase positive and Gram- negative staining tests (13).

3.2. Electrophoresis

The PCR products were separated on 1 % agarose gels (Cinna gen, Iran) in TBE 1X (Tris/borate/EDTA) buffer. Bands were visualized under UV gel documentation and photographed. Ethidium bromide (Merck, Germany) as a stain has been added to the agarose gel during preparation to give a concentration of 0.2μl/mL.

3.3. Statistical Analysis

We used absolute and relative frequency to present descriptive statistics. Chi square test as well as the Fisher exact test (when necessary) used to explain analytical statistics. Data were analyzed by SPSS version 16.0.

4. Results

DNA extracted from 63 (33%) H. pylori positive culture specimens biopsies obtained from 25 males and 38 females of whom; 46 (73%) had chronic active gastritis, 6 (9.5%) ulcerative disease includes PU & DU and 11 (17.5%) non-ulcer dyspepsia (NUD). The prevalence of the babA2 was 34 (54%) (Figure 1) and the prevalence of iceA1 and iceA2 genes were 14 (22.2%) (Figure 2) and 34 (54%) (Figure 3), respectively. In this study, the iceA2 amplicons yielded both the 229bp and 334bp fragments (Figure 3).

PCR Products Gel Electrophoresis for babA2 Gene
M:1000bp DNA Marker (Fermentas, lituania), line1: negative control, lines 2,3,5: babA2 gene positive samples, line 4: babA2 gene negative sample
PCR Gel Electerophoresis for iceA1 Gene
PCR Gel Electerophoresis for iceA1 Gene
PCR Products Gel Electrophoresis for iceA2 Gene.
Line 1: negative control. Lines 2, 5, 6 and 7: iceA2 negative samples. Lines 3, 8 and 9: iceA2 positive samples (229bp fragment). Line 4: iceA2 positive sample (334bp fragment)



  • 1.

    Atherton JC. The pathogenesis of Helicobacter pylori-induced gastro-duodenal diseases. Annu Rev Pathol. 2006;1:63-96. [PubMed ID: 18039108].

  • 2.

    Abdollahi H, Zahedi MJ, Moghadam SD, Abasi MH. A study of rdxA gene deletion in metronidazole resistant and sensitive Helicobacter pylori isolates in Kerman, Iran. Jundishapur J Microbiol. 2012;4(2):.

  • 3.

    Mohammadi M, Doroud D, Mohajerani N, Massarrat S. Helicobacter pylori antibiotic resistance in Iran. World J Gastroenterol. 2005;11(38):6009.

  • 4.

    Fujioka T, Honda S, Tokieda M. Helicobacter pylori infection and gastric carcinoma in animal models. J Gastroenterol Hepatol. 2000;15 Suppl:D55-9. [PubMed ID: 10759221].

  • 5.

    Kusters JG, van Vliet AH, Kuipers EJ. Pathogenesis of Helicobacter pylori infection. Clin Microbiol Rev. 2006;19(3):449-90. [PubMed ID: 16847081].

  • 6.

    Rudi J, Kolb C, Maiwald M, Kuck D, Sieg A, Galle PR, et al. Diversity of Helicobacter pylori vacA and cagA genes and relationship to VacA and CagA protein expression, cytotoxin production, and associated diseases. J Clin Microbiol. 1998;36(4):944-8. [PubMed ID: 9542913].

  • 7.

    Zhang ZW, Dorrell N, Wren BW, Farthingt MJ. Helicobacter pylori adherence to gastric epithelial cells: a role for non-adhesin virulence genes. J Med Microbiol. 2002;51(6):495-502. [PubMed ID: 12018657].

  • 8.

    Hennig EE, Mernaugh R, Edl J, Cao P, Cover TL. Heterogeneity among Helicobacter pylori strains in expression of the outer membrane protein BabA. Infect Immun. 2004;72(6):3429-35. [PubMed ID: 15155649].

  • 9.

    Magalhaes A, Reis CA. Helicobacter pylori adhesion to gastric epithelial cells is mediated by glycan receptors. Braz J Med Biol Res. 2010;43(7):611-8. [PubMed ID: 20521012].

  • 10.

    Yamaoka Y, Kodama T, Gutierrez O, Kim JG, Kashima K, Graham DY. Relationship between Helicobacter pylori iceA, cagA, and vacA status and clinical outcome: studies in four different countries. J Clin Microbiol. 1999;37(7):2274-9. [PubMed ID: 10364597].

  • 11.

    Ciftci IH, Uslan I, Dilek FH, Asik G, Ozgur MA, Dilek ON. [Investigation of Helicobacter pylori iceA1 and iceA2 genes in patients with chronic gastritis and gastric cancer]. Mikrobiyol Bul. 2011;45(2):228-33. [PubMed ID: 21644065].

  • 12.

    Xu Q, Morgan RD, Roberts RJ, Xu SY, van Doorn LJ, Donahue JP, et al. Functional analysis of iceA1, a CATG-recognizing restriction endonuclease gene in Helicobacter pylori. Nucleic Acids Res. 2002;30(17):3839-47. [PubMed ID: 12202769].

  • 13.

    Mishra KK, Srivastava S, Garg A, Ayyagari A. Antibiotic susceptibility of Helicobacter pylori clinical isolates: comparative evaluation of disk-diffusion and E-test methods. Curr Microbiol. 2006;53(4):329-34. [PubMed ID: 16972131].

  • 14.

    Van Doorn LJ, Figueiredo C, Megraud F, Pena S, Midolo P, Queiroz DM, et al. Geographic distribution of vacA allelic types of Helicobacter pylori. Gastroenterology. 1999;116(4):823-30. [PubMed ID: 10092304].

  • 15.

    Ghasemian Safaei H, Havaei S, Tavakkoli H, Eshaghei M, Navabakbar F, Salehei R. Relation of bab A2 genotype of Helicobacter pylori infection with chronic active gastritis, duodenal ulcer and non-cardia active gastritis in Alzahra hospital Isfahan, Iran. Jundishapur J Microbiol. 2010;3(3):93-8.

  • 16.

    Talebi bezmin abadi A, Mohabati mobarez A, Taghvaei T. An investigation of the prevalence of iceA genotypes in Helicobacter pylori strains isolated from peptic ulcer patients in Sari (2008). Arak Univ Medl Sci J. 2010;13(3):84-90.

  • 17.

    Gerhard M, Lehn N, Neumayer N, Boren T, Rad R, Schepp W, et al. Clinical relevance of the Helicobacter pylori gene for blood-group antigen-binding adhesin. Proc Natl Acad Sci U S A. 1999;96(22):12778-83. [PubMed ID: 10535999].

  • 18.

    Oliveira AG, Santos A, Guerra JB, Rocha GA, Rocha AM, Oliveira CA, et al. babA2- and cagA-positive Helicobacter pylori strains are associated with duodenal ulcer and gastric carcinoma in Brazil. J Clin Microbiol. 2003;41(8):3964-6. [PubMed ID: 12904430].

  • 19.

    Yamaoka Y, Souchek J, Odenbreit S, Haas R, Arnqvist A, Boren T, et al. Discrimination between cases of duodenal ulcer and gastritis on the basis of putative virulence factors of Helicobacter pylori. J Clin Microbiol. 2002;40(6):2244-6. [PubMed ID: 12037098].

  • 20.

    Mizushima T, Sugiyama T, Komatsu Y, Ishizuka J, Kato M, Asaka M. Clinical relevance of the babA2 genotype of Helicobacter pylori in Japanese clinical isolates. J Clin Microbiol. 2001;39(7):2463-5. [PubMed ID: 11427555].

  • 21.

    Peek RJ, Thompson SA, Donahue JP, Tham KT, Atherton JC, Blaser MJ, et al. Adherence to gastric epithelial cells induces expression of a Helicobacter pylori gene, iceA, that is associated with clinical outcome. Proc Assoc Am Physicians. 1998;110(6):531-44. [PubMed ID: 9824536].

  • 22.

    Smith SI, Kirsch C, Oyedeji KS, Arigbabu AO, Coker AO, Bayerdoffer E, et al. Prevalence of Helicobacter pylori vacA, cagA and iceA genotypes in Nigerian patients with duodenal ulcer disease. J Med Microbiol. 2002;51(10):851-4. [PubMed ID: 12435064].

  • 23.

    Ito Y, Azuma T, Ito S, Suto H, Miyaji H, Yamazaki Y, et al. Sequence analysis and clinical significance of the iceA gene from Helicobacter pylori strains in Japan. J Clin Microbiol. 2000;38(2):483-8. [PubMed ID: 10655332].