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تاثير مستخلص الكركم الخام على التعبير الجيني لمورثة البروتين المنظم للغشاء الحيوي لجرثومة Streptococcus mutans المعزولة من التجويف الفمي == Effect of Curcumine Crude Extract On The Gene Expression of Biofilm Regulatory Protein A (Brp A) In Streptococcus Mutans Isolated From Oral Cavity
Author name:
نزار احمد شهاب
Supervisor name:
عدنان حمد عبيد الحمداني | عدنان حمد عبيدالحمداني
General topic:
Medicine
Specific topic:
Microbiology
Degree:
Master
University:
University of Al-Qadisiyah - Faculty Of Medicine
Language:
English
University location:
Qadisiyah
First pages:
19T1104 - p.pdf
Abstract:
جمعت 100 مسحة فموية من مرضى تسوس الاسنان وامراض ما حول الاسنان الذين تراوحت اعمارهم ما بين 15 - 65 سنة ومن كلا الجنسين والذين راجعوا المركز التخصصي لطب الاسنان في الديوانية للمدة من كانون الاول 2012 الى اذار 2013 لغرض قياس التعبير الجيني لمورثة البروتين ا | A total of 100 oral swabs were collected from patients suffering from dental caries and priodontal infections, ranging from 15 - 65 years old of both genders whom admitted to the Al - Diwaniyah Teaching Special Center of Dentistry within the period from December 2012 to March 2013, in order to quantify the gene expression of biofim regulatory protein (Brp A) in Streptococcus mutans isolates by using the Real Time polymerase chain reaction. According to morphological, cultural characteristics and biochemical tests by using VITEK - 2 system, 34/84(40.47%) isolates of S.mutans were isolated and identified among streptococcus species and the isolation percentages were 26/50(52%) in males and in females were 24/50(48%), while the age group (15 - 25 years old) was the most affected than the other groups. The statistical analysis showed no significant differences among the interaction between gender and age groups. Molecular detection of 16s rRNA gene was amplified to confirm the diagnosis of S.mutans isolates, where all isolates gave an amplicon size (151) bp, in addition to, the DNA sequencing was done in order to identify the nitrogen bases sequences and phylogeny of local isolates in comparison with the global isolates (Spain, Japan, Korea, USA, Canada, India), results revealed that the local isolates of S.mutans had a similarity percent (90%) with Spain isolates.Furthermore, the susceptibility of isolates to a variety of antibiotics have been investigated. It has been found that isolates had multiresistance to erythromycin(88.2%),ampicillin(76.4%),amoxiclave(70.5%),tetracycline(61.7%), nalidixic acid and amoxicillin (58.8%), chloramphenicol (55.8%). The statistical analysis showed a significant differences among tested antibiotics. Biofilm formation was investigated phenotypically by using tissue culture plate with crystal violet staining for resistant S.mutans isolates, the results revealed that all isolates were high biofilm former, furthermore, the effect of ethanolic extraction (50%) of curcumine in growth inhibition and biofilm reduction of S. mutans was assayed. The results showed an effect of curcumine in comparison with chlorhexidine(2%) as a control.The gene expression of biofilm regulatory protein A(Brp A) was quantified in comparison with housekeeping gene 16s rRNA as a reference gene of biofilm S.mutans by using reverse transcriptase polymerase chain reaction (RT - PCR). The results revealed that the expression (relative&absolute) of Brp A gene is decreased in the test treatment (S.mutans with 1% glucose+ 50% curcumine) in comparison with control (S.mutans grown in 1% glucose only).In conclusion, the present study established the role of Brp A gene in biofilm formation
