الفهرس 
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Abstract
The present investigation was carried out to study the effect of both photodynamic treatments (using He-Ne laser and Toluidine blue O) to medication treatment (using Azithromycin) on bacterial growth. It was aimed also to assess the effect of both methods of treatment on H.pylori genome using RAPD-PCR. The literature was reviewed including laser physics, laser tissue interaction, photodynamic therapy (PDT), Helicobacter pylor, and molecular studies on H.pylori. Biopsies were taken from 10 duodenal ulcer male patients. Gram-stain and biochemical used for identification of H.pylori. Helicobacter pylori ATCC 43504 used for quality control. He-Ne (632.8 nm) laser was used for irradiation. The photosensitizers used was toluidine blue O (TBO). The energy density of He-Ne laser used for killing 93 % of bacteria was 210 J/Cm2 with exposure time 5 minutes. The susceptibility of Helicobacter pylori isolates and Helicobacter pylori ATCC 43504 to metronidazole and Azithromycin were examined by ε-test. DNA was extracted from H.pylori groups (control, laser treated and antibiotic treated groups). RAPD fingerprint profiles were carried out to assess the effect of PDT and medical treatment using nine random 10-mer primers on genomic DNA. RAPD products were analyzed by 1.5% (W/V) agarose gel electrophoreses. The resulting data were statistically analyzed to investigate similarity index and commonality among all investigated groups. The results of the present investigation have been discussed compared to that done by others. And the reached results were summarized in the following: Nine out of the ten duodenal ulcer patients were H.pylori positive. The energy density 210 J/Cm2 from He-Ne laser with TBO photosensitizer with concentration 100g/ml used to kill 93% of Helicobacter pylori isolates and Helicobacter pylori ATCC 43504. Helicobacter pylori isolates and Helicobacter pylori ATCC 43504 were sensitive to Azithromycin and resistant to Metronidazole.The Azithromycin MICs for Helicobacter pylori isolates and Helicobacter pylori ATCC were ranged between 0.25-0.5 μg/ml The number of RAPD produced fragments were 44 distributed as 2,8,3,13,10&8 fragments with primer OPA.1, OPA.3, OPA.7, OPA.9, OPA.17& OPA.19 respectively. Random primers OPA5, OPA11 and OPA20 failed to generate any product. The molecular sizes of fragments ranged from 204 bp to 1473 bp. Bands (No.12, 416 bp & No.15, 268 bp) seemed to be characteristic to H.pylori species where, they detected in all investigated groups by using six primers and five primers respectively. These common bands may represent genetic marker for H.pylori species. The mean value of similarity index between H.pylori groups and H.pylori ATCC groups (Hc/Ac, Ha/Aa & HL/AL) using the six primers was 0.88, while the mean value of band sharing percentage between the same groups was 79 %.The mean value of similarity index between control groups and laser treated groups (Hc/HL, & Ac/AL) using the six primers was 0.87, while the mean value of band sharing percentage between the same groups was 77.5 %. The mean value of similarity index between control groups and antibiotic treated groups (Hc/Ha, & Ac/Aa) using the six primers was 0.92, while the mean value of band sharing percentage between the same groups was 85 %.The results of this study show that, nine RAPD primers were used to screen genetic polymorphism in DNA of different H.pylori groups. Six of them produced RAPD products while three failed to generate any product. Two bands (No. 12, 416 bp & no.15, 268 bp) may represent genetic marker for H.pylori species or higher taxonomic position on family or order levels but further techniques should be applied to verify the identity of these markers. The resulting data showed that, although the overall genetic differences between control groups and laser treated groups was higher than that between control groups and azithromycin treated groups yet it still law genetic variability. These indicate that, the primary site of attack which was the main cause of cell death of both PDT and azithromycin was not DNA. The main cause of cell death of PDT using TBO as a photosensitizer was mainly cell wall and cytoplasmic membrane.