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Paper Information

Journal:   IRANIAN JOURNAL OF PUBLIC HEALTH   AUGUST 2014 , Volume 43 , Number SUPPLEMENT 2; Page(s) 22 To 22.
 
Paper: 

COMPARISON OF CAGE AND BABA VIRULENCE GENES EXPRESSION IN SPIRAL AND COCCOID FORMS OF HELICOBACTER PYLORI

 
 
Author(s):  POURSINA FARKHONDEH, FAGHRI JAMSHID, GHASEMIAN SAFAEI HAJIEH*
 
* DEPARTMENT OF MICROBIOLOGY, FACULTY OF MEDICINE, ISFAHAN UNIVERSITY OF MEDIAL SCIENCES, ISFAHAN, IRAN
 
Abstract: 

Background: Under stressful environment, a spiral form of H. pylori is transformed into a coccoid form. The coccoid formation is very important problem in recurrence of disease and antibiotic treatment failure in gastritis pateints. Our objective of this study was to determine the morphological transformation-process from spiral to a coccoid form under Amoxicillin stress and also to evaluate and comparison of the virulence genes (cagE and babA) expression in spiral and coccoid forms of H. pylori.
Methods: The standard ATCC 26695 H. pylori strain and 9 clinical isolates were used in this study. The existence of interest genes were detected by conventional PCR method. Minimal inhibitory concentration (MIC) of amoxicillin was determined using by E-test method. All of ten isolates were cultured under MIC, 1/2, 1/4 and 1/8 MIC concentrations of amoxicillin. The induction of coccoid forms was examined periodically by gram staining and light microscopy. Then, the viability of induced coccoids was evaluated by flow cytometry method. The quantitative mRNA expression levels of cagE and babA genes were detected and compared in spiral and coccoid forms of H. pylori by real-time PCR method.
Results: The MIC and sub MIC (1/2) of amoxicillin could be induce the highest rate (100% and 98% respectively) of coccoids H. pylori after 144 h. Flow cytometry analysis revealed that the MIC and sub MIC (1/2 MIC) concentration of amoxicillin induced the viable coccoids (
>80% and 4% respectively). Quantitative RT-real-time PCR showed that the expression of cagE and babA in coccoids were lower (1.9 and 6.1 fold) than spirals, however, the difference between cagE expression was not significant in two forms.
Conclusion: Amoxicillin in sub MIC concentrations is a potent stressor for inducing the viable but non-culturable (VBNC) coccoid form of H. pylori. Of interest was the expression of two important virulence genes (cagE and babA) in coccoids, however with lower rate than spirals. In conclusion, we have concluded that the coccoid cells may have a role inbacteria lresistance, persistency and disease recurrence.

 
Keyword(s): HELICOBACTER PYLORI, GENE EXPERSSION, STRUCTURAL GENE VIRULENCE GENE, SPIRAL, COCCOID FORM
 
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