PHD theses : Medical Laboratory Science
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Item Phenotypic and Genotypic Detection of Streptococcus pneumoniae associated with Pulmonary Tuberculosis in Patients attending Kasala Teaching Hospital(2017) Magdi Babiker Omer MohammedABSTRACT Background: Streptococcus pneumonia (S. pneumoniae) is the most frequent etiologic agent of bacterial pneumonia which develops when encapsulated "virulent" S. pneumoniae is inhaled into the alveoli of susceptible hosts. Objective: This study aimed to determine the phenotypic and genotypic characterization of S. pneumoniae associated with pulmonary tuberculosis and also to evaluate the effect of antituberculosis treatment of S. pneumoniaein patients attending Kasala Teaching Hospital Materials and methods: One hundred and fifty sputum specimens were collected from patients suffering from pulmonary tuberculosis. S. pneumoniae was isolated by culture on chocolate agar medium and biochemical tests were employed for identification. Kirby Bauer method was used to detect susceptibility of isolates to antibacterial agents. At the same time part of the sample used to detect the genotype of S. pneumoniae by PCR technique to detect cpsA gene . Results: Out of the 150 sputum specimens 21(14%) specimens were found culture positive for S. pneumoniae. The distribution of isolated S. pneumoniae among gender was 44 females were (7/15.9%) and 106 males was (14/13.2%). One hundred and one of the patients investigated were residents of Kasala City and 9 (42.9%) of them were harboring S. pneumoniae; while 49 of the patients studied were residents outside Kasala City and 12 (57.1%) of them were harboring S. pneumoniae. According to age groups, most of S. pneumoniae isolates were in the age range 51-60 years (frequency rate 28.6%). All 150 samples were extract the genes (cpsA) to investigated with the PCR and the result was 27(18.0%) which is greater than the isolate by culture method 21(14.0%). The antibiotic susceptibility pattern revealed that all S. pneumonia isolates (21/100%) were sensitive to vancomycin, penicillin, tobramycin, and oxacillin. Also, S. pneumonia isolates were more sensitive to rifampicin (19/90.5%) and pyrazinamide (19/90.5%); followed by streptomycin (17/81%), ethambutol (16/76%), and isoniazid (14/17%). Conclusion: Some S.pneumonia was multi-drug resistant in which 4 of S. pneumonia isolate show multi drug resistance 3 isolated restist (isoniazid, E thambutol, and stryptomycin ) and one isolate resist(isoniazid, E thambutol, Rifampicin and stryptomycin ) . The frequency rate of S. pneumoniae infection among females was higher than that among males. Also PCR technique was more sensitive than culture method. الخلاصة الخلفية:-تعتبر بكتربا المكورات العقدية الرئوية من اكثر المسببات للاتهاب الرئوي وذلك لوجود عوامل امراضية ، والتي تنتقل عبر الاكياس التنفسية داخل الرئتين مما يسبب مشكلة اساسية للاشخاص المصابين بالسل الرئوي . الاهداف:-هذه الدراسة صممت للبحث عن الخصائص الجينية والشكلية لبكتريا المكورات العقدية الرئوية التي تتواجد في الاشخاص المصابين بمرض السل الرئوي مع تقييم تاثير علاج السل الرئوي وتاثيره علي هذه المكورات المواد وطرق البحث:-150 عينة من القشع جمعت من المرضى الذين يعانون من التهاب السل الرئوي وعزلت المكورات العقدية الرئوية في وسط عزل غني بالدم وتم استخلاص الحمض النووي مباشرة من القشع لمعرفة الجين عن طريق تقنية سلسلة التفاعلات البلمرية لمقارنته بطريقة الزراعة التقليدية النتائج:-في 150 عينة من القشع كانت نسبة العزل للمكورات العقدية الرئوية عن طريق التزريع 21 (14%) وكان توزيعها 44 عينة عند الاناث بنسبة عزل (7/15.9%) و106عينة عند الذكور بنسبة عزل (14/13.2%). 101 عينة من مرضى السل كانوا من سكان مدينة كسلا بنسبة عزل (9/42.9%) للمكورات . والذين من خارج مدينة كسلا كانت بنسبة عزل (12/57.1%)، وبالرجوع للفئات العمرية كانت الاعمار 51-60 سنة اكثر اصابة (28.6%) بالمكورات. كل العينات تم فحصها لاستخلاص الجين عن طريق تقنية سلسلة التفاعلات البلمرية وكانت النتائج 27 عينة ايجابية بنسبة 18% وهذه النتيجة تعتبر ادق من نتائج التزريع والتي وصلت نسبتها الى (21/14%). أما بالنسبة لاختبارات الحساسية للمضادات الحيوية فقد كانت حساسة بنسبة عالية للفانكومايسين والبنسلين والأكثاسلين والتبرامايسين. وبالنسبة لاختبارات الحساسية للمضادات الحيوية الخاصة لجرثومة السل الرئوي فكانت المكورات العقدية الرئوي حساسة بنسبة عالية للريفامبسين 19(90.5%) والبارازاماين 19(90.5%) ثم الاستربتومايسين 17(81%) ثم ايثامبيتول 16(76%) ثم ايزومازايت 14(67%) . الاستنتاج : تم عزل المكورات العقدية الرئوية من المصابين بمرض السل الرئوي رغما عن تعاطيهم للعلاج الخاص بالسل الرئوي نتيجة لمقاومتها لهذا العلاج . الاناث اكثر عرضة للاصابة بالمكورات العقدية الرئوية . وطريقة الفحص بنظام تقنية سلسلة التفاعلات البلمرية اكثر دقة في العزل.Item The Effect of Transfer Factor and BCG as Immunoprotective in Mice infected with Mycobacterium tuberculosis(AL-Neelain University, 2017) Jamal Baid Salim MohammedABSTRACT Background: Tuberculosis, caused by Mycobacterium tuberculosis was one of the most serious diseases in Sudan and elsewhere. The disease was quite prevalent in Eastern Sudan. Objective The aim of this experimental study is to determine the protective efficacy of transfer factor (TF) as immunoprotective for mice in comparison to BCG. Materials and Methods To evaluate the immunoprotection of experimental mice by several parameters including; survival of challenged mice, mortality rate and delayed type hypersensitivity A total number of 102 mice were used. Experiment(1) n=10 for each group A and B for the susceptibility and resistance of the strains of mice by immunization of mice with BCG for 21 days and testing by tuberculin skin test (TST). experiment(2) preparation of T.F. n = 10. Experiment(3) contains three groups of mice of 10 mice n = 10 each . The first group(A) were immunized with BCG (0.2ml) first dose (I.P) for 21 days. The second group (B) were immunized with BCG (I.P) as first dose for 15 days and boosting dose (second dose) for another 15 days. The third group(C) were not immunized with BCG. All the three groups were challenged with (0.5ml)virulent M.TB. Experiment(4) Three groups of mice were used n= 12 for each group (36mice), the groups were T1,T2 andT3 .The mice of group T1and T2 were given murine transfer factor(mTF) which prepared from the spleen of infected mice and 24 hours later all the mice of groups T1and T3 were challenged with virulent M. tuberculosis H37Rv strain (American Type Culture Collection, ATCC 35718)M.TB.107(CFU).Group T2 was not challenged with MTB . Experiment(5) n=6 for the study of humoral response by immunization of mice with immune serum and challenged with M. tuberculosis H37Rv strain. Results: After three weeks of observations the mice of experiment(1) were tested for tuberculin skin test and the result were positive. Following the results of experiment(2), survival mice in group (A) were 50%, group (B) 70% and group(C) 0%.The mortality rates for (A) 50%, (B) 30% and (C) 100%.Experiment(3) results of Effectiveness determination of murine transfer factor (mTF) of the experiment as the rate of challenged survival mice wereT1(83.3%), T2(100%) and T3 was (0 %) and mortality rates wereT1(16.7%), T2 (0 %)and T3(100%). Tuberculin skin test reaction for the BCG immunized mice were positive, hence the mice strain of BALB/c were susceptible and therefore T.F can be prepared, while the strain of Swiss white mice were resistant for BCG. The immunopotency and protective efficacy of BCG first dose and boosting dose were (50%) and (70%) respectively while effectiveness determination of TF as protective efficacy was (83.3%). Humoral immunity response against M.TB .showed negative reaction hence mortality rate was 100%. Conclusions: Transfer factor (TF) was shown to be capable of transferring antigen-specific cell-mediated immunity (CMI) to T lymphocytes and therefore TF can successfully be used as treatment for M.TB. and specially for MDR-TB and supplementary for conventional chemotherapy, in which CMI plays a relevant role in protection and control of the disease, such as intracellular bacterial diseases (tuberculosis, leprosy) and parasite infections (leishmaniasis, toxoplasmosis).The incidence of TB is high inspite of primary vaccination in neonatal period and therefore requires consideration for repeated immunization of BCG. Research concerning TF are going on to discover secrets of TF in protection from diseases, increasing and transferring of immunity in patients.