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التحري عن التشوهات الكروموسومية وطفرات المورث FLT3/ITD في مرضى ابيضاض الدم النخاعي الحاد Screening For Chromosomal Aberrations And Gene Mutations FLT3/ITD In Patients With Acute Myeloid Leukemia

اسم المؤلف: سمارة كاظم محمد
اسم المشرف: عبد الحسين مويت الفيصل
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد - معهد الهندسة الوراثية والتقنيات الاحيائية - قسم التقانة الاحيائية
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: صممت الدراسة الحالية من اجل تسليط الضوء على تاثيرات مرض ابيضاض الدم ((AMLفي بعض معايير الوراثة الخلوية والجزيئية الحاصلة في كريات الدم البيضاء في ثلاثة حالات للمرضى اشتملت على مرحلة ما قبل العلاج, في اثناء العلاج وبعد العلاج الكيميائي. كما شملت الدراسة ا | The present study was designed to shed light on the cytogenetic and molecular effects caused by acute myeloid leukemia (AML) pathogenesis in three stages before, during and after treatment with chemotherapy (in vitro) in lymphocytes. It was also aimed to investigate chromosomal aberrations, micronucleus formation and FLT3/ITD point mutations in CN - AML region 100 - 300 bp compared with healthy control. The study was comprised of forty - seven AML Iraqi patients their ages ranged between 2.5 - 81 years. It included twenty females and twenty seven males compared with twenty - six healthy control. The following results were obtained : AML is most common leukemia in adults and uncommon in children. It was found that 46.8% patients suffer from AML, who were under 15 years old, while 19.15% patients their age ranged between 16 - 30 years; 19.15% of patients their age were more than 45 years and 14.9% of patients their age ranged between 31 - 45 years of the total AML patients. AML is more common in males than females. The percentages of females and males were 42.5% and 57.5% respectively representing 1.35 : 1.00 male : female ratio. Two AML patients 5.3% have diploid cells when examined under light microscope. The highest mitotic index was 7.498±1.7 occurred in patients before taking treatment while recorded 6.784±4.5 during treatment and 7.000±2.5 after treatment. Nuclear anomalies such as nucleoplasmic bridges and nuclear buds were observed in AML patients, Micronucleus mean values recorded 0.033±0.018, 0.020±0.00015 and 0.036±0.01for AML patients before, during and after treatment respectively when compared with the control which recorded 0.002±0.00002. Nuclear division index (NDI) means before, during and after treatment of AML patients were 1.658±0.2, 1.000±0.022 and 1.424±0.19 respectively. Significant differences occurred among the three groups when compared with the control group (1.282±0.09). Extracted DNA from 30 AML patients was amplified by PCR to obtain FLT3/ITD gene from exon 11 to intron 12 and ten of them sent for sequencing. Two patients showed larger bands (470 and 460) bp when compared with wild type (330) bp. Among six patients, three displayed point mutations of deletion and substitution while others were normal since no mutations were detected. The percentages of mutation types were substitution 57.78% and deletion 22.2%. The effect of mutations was missense mutation (55.54%), deletion (22.22%) and nonsense (22.22%). It has been concluded from the current work that AML is more common in adult males, patients suffer from AML exhibited high MI, NDI, MN and point mutations are present in such patients including deletion and substitution causing missense, nonsense and deletion.

دراسة جزيئية عن جين المقاومة mecA في بكتريا العنقودية الذهبية المقاومة للمشيسلين والمعزولة من بعض مستشفيات بغداد Molecular Study For Detection of Meca Gene In Methicillin - Resistant Staphylococcus Aureus Isolated From Some Hospital In Baghdad City

اسم المؤلف: لمى ياسين موسى
اسم المشرف: محمد ابراهيم نادر
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد - معهد الهندسة الوراثية والتقنيات الاحيائية - قسم التقانة الاحيائية
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: Two hundred and thirty clinical specimens were collected from two different hospitals in Baghdad during the period (December 2012 to April 2013). These specimens were collected from visitors, hospitalized patients and the health care workers in these hospitals. The specimens included nasal swab, wound swab, burn swab, abscess and pus, sputum, ear swab, urine and blood culture diagnostic results show that 150 out of the (230) specimens gave positive bacterial cultures and (100) isolates are characterized as Staphylococcus aureus depending on the cultural and biochemical examinations.the coagulase test was performed and the results showed that from total 150 isolates of Staphylococci, 100 isolates (61%) were coagulase - positive (COPS), while only 50 isolates (39%) were coagulase negative (CONS). In addition, the distribution of methicillin resistance among Staphylococcus spp. was investigated.The use of the antibiotic susceptibility profile for these isolates was examined against methicillin resistance. Using disk diffusion method revealed that (13) isolates were proved to be methicillin resistant Staphylococcus aureus (MRSA), While (87) isolates of S. aureus showed sensitivity to methicillin (MSSA) and there was no intermediate resistance among these isolates.The ability of MRSA isolates to produce some virulence factors were investigated and the results showed that MRSA isolates produce many enzymes and toxins that contributed in their virulence such as protease, urease, dnase and gelatinase, and also produce a beta types of haemolysins.The ability to produce slime layer by MRSA isolates was also investigated and the results showed that all isolates of MRSA were produced slime layer when theytested by Congo red agar method and the results showed that all of MRSA isolates produced strong slime layer.One of the aims of this study was to adopted a accurate diagnostic method to detect S.aureus by its genetic material contents through extracting of DNA and gel electrophoresis of the PCR product for the specific gene.Detection of methicilline - resistance gene represented by A confirmatory test was carried out for the selected isolates using Polymerase chain reaction (PCR) technique for further characterization up to the species level by the amplification of (mecA) gene.This is Staphylococcus aureus specific gene that encodes the extra Penicillin Binding Protein, which is unique to methicillin - resistant staphylococci. All the (13) positive isolates by disk diffusion test are found to be positive for the presence of (mecA) gene as their agarose gel revealed the presence of DNA band of mecA gene with a molecular size about (200 bp.).Results of the detecting (femB) gene showed that it was positive in all of MRSA isolates as they appear to have a band with a molecular size of about (651 bp). The genetic determinants of methicillin resistance mecA and femB genes were amplified using multiplex PCR technique in order to identify methicillin resistant (mecA+) and susceptible (lacking mecA) staphylococci and to differentiate S. aureus (femB+) from coagulase negative staphylococci (lacking femB). All of the S. aureus isolates (100%) were found to harbor femA gene, it is species specific marker for S. aureus.