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التتابع الجيني لطفيليات Entamoeba histolytica, E. dispar and E. Moshkoviskii في محافظة القادسية == Phylogenetic Sequeincing For Spesies Entamoeba Histolytica, E. Dispar, E. Moshkoviskii In Al - Qadisiya Province
Author name:
هبة رياض جميل العبودي
Supervisor name:
هادي مدلول حمزة الميالي
General topic:
Biology
Specific topic:
Microbiology - Parasites
Degree:
Doctorate
University:
University of Al-Qadisiyah - Faculty Of Education - Department Of Biology
Language:
Arabic
University location:
Qadisiyah
First pages:
24T2949 - p.pdf
Abstract:
اجريت هذه الدراسة للمدة من بداية اذار 2014 والى نهاية ايلول 2014 وشملت على مراجعين مستشفى الديوانية العام ومستشفى النسائية والاطفال في محافظة القادسية ومختبرات التحليل الاهلية في المحافظة, وقد تم فحص679 عينة براز باستخدام طريقة المسحة المباشرة لمختلف الفئ | This study has been conducted for the period from the first of March 2014 until September 2014 and included the auditors to Diwaniya General Hospital and Women's Hospital and children in the province of Diwaniya and civil analytical laboratories in the province. A sample of 679 of auditors' stool have been examined by using the direct swab from different age groups and both sexes. The study shows that 61.26% of people have been infected by screened amoeba parasite. Real time PCR was used in this study to diagnose Amoeba species in 200 samples. Result shows that Entamoeba histolytica had highest percentage of incident followed by Entamoeba dispar then Entamoeba moshkoviskii, which were 74 %, 26 %, and 7 % respectively.Statistical level of ?=0.05presents significant differences between then. Percentage of positive molecular test samples was higher than the negative ones; they were 88% and 12%. Molecular diagnoses were used in this study. E. histolytica had highest individual incident followed by E.disbar then E. moshkoviskii : 56% 10%, and 1% respectively. Using significant level of 0.05, significant differences were appeared. Rates of mixed infections of E. histolytica + E. dispar followed by E. moshkoviskii + histolytica then E. moshkoviskii + E. dispar, which were 14%, 4%, and 2% respectively. Statistics shows significant differences between them at ? =0.05. Symptoms of individual incident of the three Amoeba sorts were observed and recorded. Patients who had E. histolytica, severed from acute diarrhea that ranged between mucous and watery texture, which its color contrast between brown and reddish brown. They had bloody diarrhea and abdominal pain that ranged from severe to moderate colic. They had also slight temperature increase and weight loss. For those who had E.disbar, abdominal pain and frequent pale yellow diarrhea that semi - aqueous texture were recorded. Symptoms of : moderate yellowish brown snotty diarrhea, abdominal pain and, weight loss were observed for those who had E. moshkoviskii. R.T.PCR (Taqman Probe) was used to isolate samples and to investigate the three pathogenic virulence genes, which are Cystiene Proteinase, Amebopore, and Gal/Gal Nac Lectin. This was done in each of Amoeba’s species using R.T.PCR (Syber Green). According to it, E. histolytica possesses Cystiene Proteinase 1, Amebopore, and Gal/Gal Nac Lectin at 86.6%, 80.35%, and 42.8% respectively. Non - significant differences were found at ? =0.05 between them. For E.disbar, result shows existence of the same pathogenic virulence genes at percentages of 65%, 45%, and 20% respectively. Result presents significant differences among percentage of genes isolation for the pathogenic virulence factors at ? =0.05. E. moshkoviskii had these three pathogenic virulence genes at percentage of 33.3% by isolating the virulence genes except Gal/Gal Nac Lactin, there were significant differences among them at ? =0.05. This study was conducted to understand the molecular characterization confirmatory of the three types of insulators parasite using traditional enzyme chain polymerize reaction as a first stage to investigate the 18s ribosomal RNA covenantal. It represents personal heritage of the three types of amoeba. Depending on the same gene, developed tree were analyzed for Amoeba’s species using method of Neighber - Joining among different groups of the stains for each species separately using MEGA6 program. Knowing convergence rates of the used stains and international isolators from NCBI, Blast program was used. Since there is no study has been done to study nitrogenous bases sequence for these three species, samples undertook molecular characterization confirmatory was registered in the national data base center (NCBI). International serial numbers were obtained and attached to the dissertation.
