التعبير الجيني للمورث Erg11 وعلاقته بالمقاومة وظاهرة رجوع النمو لبعض عزلات خميرة المبيضات البيضاء المختبرة تجاه مضادات الازول الفطرية == Gene Expression of Erg11 And It’s Relation With Resistance And Trailing Growth Phenomena of Some Isolates of Candida Albicans Tested Against Azole Antifungals
Author name:
حيدر عبد الحسين عباس
Supervisor name:
عدنان حمد عبيد الحمداني
General topic:
Biology
Specific topic:
Microbiology
Degree:
Master
University:
University of Al-Qadisiyah - Faculty Of Education - Department Of Biology
Language:
Arabic
University location:
Qadisiyah
First pages:
24T2881 - p.pdf
Abstract:
هدفت الدراسة الحالية الى الكشف عن دور المورث Erg11 والتعبير الجيني النسبي له في مقاومة خميرة المبيضات البيضاء C.albicans وعلاقته بنشوء عزلات مقاومة او ظاهرة رجوع النمو Trailing growth لهذه الخميرة عند اختبار حساسيتها الدوائية تجاه بعض مضادات الازول (الفلو | The current study aimed to detect the role of Erg11 gene and its relative expression in the resistance of Candida albicans, and its relationship with the emergence of resistace or Trailing growth isolates when tested its susceptibility against azoles agenst (fluconazole, ketoconazole). A total of 120 specimens were collected from patients of both gender with different ages whom infected with oral thrush, uro - genital and Gastrointestinal candidiasis and admitted the General Teaching Hospital and materinaty and children Teaching Hospital in AL - Diwaniyah city during the period from December/2012 to February / 2013. The isolation and identification of yeast isolates were done based on their morphological, Cultural and biochemical tests in addition to use the confirmatory systems such as the growth on Chrom Agar candida, Api Candida and Tobacco Agar medium to distinguish C.albicans from other yeast spp. Also, some virulence factors of C.albicans such germ tube formation, chlamydospores production, growth at of 45 c and resistance to cycloheximide were detected. The antibiotics sensitivity test was used in two method (disk diffusion & dilution) to determine the sensitive, trailing growth, resistance isolates and minimal inhibitory concentration (MICs) toward azole agents. The qRT - PCR was used to detect the relative quantitative of Erg11 gene expression and its relationship with resistance to azole and emergence of trailing growth phenomena in the presence of Fluconazoles and ketoconazole. The isolation and identification results revealed that the isolation percent of C.albicans was 47.05 % incomparison with other Candida species that were 15.3%, 11.76 %, 8.23 % and 5.9 % for C.krusi, C.dubliniensis, C.tropicalis, C. parapsilasis and C.glabrata , respectively. The antibiotic susceptibity tests of C.albicans showed the presence of resistance precent (38.89%) to fluconazole and the most of sensitive isolates revealed a trailing growth phenomena in the zone of growth inhibition of floconazole in a ratio 55.56%. While the resistance percent to ketoconazole was 27.78 % and the sensitive isolates showed the trailing growth in a ratio (38.89%). The statistical analysis showed a significant differences (P <0.05) among tested treatments. The MICs values of fluconazole against C.albicans isolates were 8, 16, and 64 Mg/ml for sensitive isolates and were 0.125 and 0.5 Mg /ml for resistance isolates and the trailing growth phenomena occurred at MICs value ? 8 Mg/ml within 24 hours of incubation while the values became ? 64 Mg/ml after 48 hours of incubation.There was a significant differences (p< 0.05) among tested treatments isolates incubated at 24 hours while there was no significant differences (P>0.05) at 48 hours of incubation. In respect with Ketoconazole, the resistance of C.albicans was increased after 48 hours of incubation at MICs values 0.125, 0.25, 0.5 and 1 Mg /ml while its was sensitive at values 4, 8 and 16 Mg /ml and the trailing growth phenomena occurred when the MICs ? 2 Mg/ml after 24 hrs of incubation, while it reached to ? 8 Mg/ml after 48 hrs of incubation. There was a significant differences (p< 0.05) among tested treatments incubated after 24 hours while there was no significant differences (P >0.05) among treatments after incubation 48 hours. The results of relative quantitative gene expression for both genes revealed a significant differences (p< 0.05) in gene expression levels through the occurrence of increased in the transcription of mRNA quantity among tested treatments.The up - regulation of Erg11 and fold change in mRNA transcription was 2.498 fold compared with 1.0 for control treatment after 24 hrs of incubation in the presence of antifungal, while the increasment was 12.606 fold after 48 hrs of incubation of isolates with antifungals.
