article information

Jundishapur Journal of Microbiology: Vol.6, issue 4; e4739
published online: June 10, 2013
article type: Research Article
received: March 7, 2012
revised: May 27, 2012
accepted: October 6, 2012
DOI: https://doi.org/10.5812/jjm.4739
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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

authors:

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. https://doi.org/10.5812/jjm.4739.

Abstract

Background:

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.

objectives:

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.

Results:

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

M:1000bp DNA Marker (Fermentas, lituania), line1: negative control, lines 2,3,5: babA2 gene positive samples, line 4: babA2 gene negative sample

Figure 1.

PCR Products Gel Electrophoresis for babA2 Gene

PCR Gel Electerophoresis for iceA1 Gene
PCR Gel Electerophoresis for iceA1 Gene
Figure 2.

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)

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)

Figure 3.

PCR Products Gel Electrophoresis for iceA2 Gene.

Acknowledgements

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