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).
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
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)
