Characterization of MycobacteriumTuberculosisIsolated from Pulmonary Tuberculosis Sudanese Patients

Abstract

VIAbstract Introduction WHO estimated that one third of the world‘s population are infected with M. tuberculosis and approximately 9.7 million new cases of tuberculosis were diagnosed globally in 2013. Sudan is considered as one of the high prevalence countries of tuberculosis. Objectives The aim of this study was to perform culture for M.tuberculosis and to determine the susceptibility of the isolates to the first line anti-tuberculosis drugs. Also the study aimed to employ the molecular techniques to determine the source and to cluster the isolated organisms and to compare them with the available international data Material and Methods This study has been conducted during the period from May 2007 to October 2013. 243 sputum specimens had been collected from patients from Port Sudan, Alobied, Khartoum and Omdurman. Sputa were treated using Petroff’s method. ZN stain was used for staining the smears and these were seen under the light microscope. L.J. medium with glycerol was used for inoculation of all specimens and incubated aerobically at 37C. Nitrate reduction test and PH7/68C catalase tests were used for identification of the growth. Sensitivity to Rifampicin, INH, Ethambutol and Streptomycin was performed using the proportion method. VIISpoligotyping and MIRU-VNTR were used to discriminate different strains of M.tuberculosis and results were compared to the international data base. Results Tuberculosis was found in different parts of Sudan and it affects all age groups. No significant difference in incidence of the disease among different tribes. 58% of the samples gave positive cultures for M. Tuberculosis in L.J. medium. The overall resistance to anti-tuberculosis drugs was 20%. Multi drug resistant strains (MDR) were 8.7% All strains were grouped into 28 different clusters-spoligotypes. A total of 31 strains have unique patterns and were considered as orphan strains. Conclusions The resistance of M.tuberculosis isolated from Sudanese patients to antituberculosis drugs was high. The MDR strains of M. tuberculosisdetected in this study was alarming (8.7%). VIII‫‪.‬‬ ‫الاصابات‬ ‫المرتفعفى‬ ‫المستوى‬ ‫ذات‬ ‫البلدان‬ ‫من‬ ‫السودان‬ ‫ویعتبر‬ ‫‪.‬‬ ‫‪2013‬‬ ‫عام‬ ‫في‬ ‫تشخیصھم‬ ‫الأھداف‬ ‫أیضا‬ ‫تھدف‬ ‫كما‬ ‫‪.‬‬ ‫المستعملة‬ ‫للأدویة‬ ‫اومتھا‬ ‫مق‬ ‫أنواع‬ ‫وتحدید‬ ‫السلیة‬ ‫المتفطرة‬ ‫لزراعة‬ ‫الدراسة‬ ‫ھذه‬ ‫تھدف‬ ‫ومقارنتھا‬ ‫السودان‬ ‫في‬ ‫المرضى‬ ‫من‬ ‫المعزولة‬ ‫الأنماط‬ ‫منشأ‬ ‫لمعرفة‬ ‫الجزیئیة‬ ‫الحیویة‬ ‫الطرق‬ ‫لاستخدام‬ ‫‪.‬‬ ‫العالم‬ ‫أجزاء‬ ‫ببقیة‬ ‫البحث‬ ‫وطرق‬ ‫العینھ‬ ‫لت‬ ‫شم‬ ‫السودان‬ ‫من‬ ‫مختلفة‬ ‫أجزاء‬ ‫في‬ ‫‪2013‬‬ ‫أكتوبر‬ ‫الى‬ ‫‪2007‬‬ ‫مایو‬ ‫من‬ ‫الفترة‬ ‫في‬ ‫الدراسة‬ ‫ھذه‬ ‫أجریت‬ ‫في‬ ‫بالسل‬ ‫مصابین‬ ‫مرضى‬ ‫من‬ ‫(‬ ‫بلغم‬ ‫)‬ ‫تفاف‬ ‫عینات‬ ‫أخذ‬ ‫تم‬ ‫حیث‬ ‫والخرطوم‬ ‫والأبیض‬ ‫بورتسودان‬ ‫من‬ ‫كل‬ ‫بورتسودان‬ ‫ومستشفى‬ ‫الأبیض‬ ‫ومستشفى‬ ‫عنجة‬ ‫أبو‬ ‫ومستشفى‬ ‫بالخرطوم‬ ‫الشعب‬ ‫بمستشفى‬ ‫السل‬ ‫عیادات‬ ‫‪.‬‬ ‫موافقتھم‬ ‫أخذ‬ ‫بعد‬ ‫تم‬ ‫ثم‬ ‫نیلسن‬ ‫زیل‬ ‫صبغة‬ ‫باستعمال‬ ‫وتجھیزھا‬ ‫تحضیرھا‬ ‫بعد‬ ‫التفاف‬ ‫لعینات‬ ‫المجھري‬ ‫الفحص‬ ‫عمل‬ ‫تم‬ ‫للتعرف‬ ‫المعزولة‬ ‫للبكتریا‬ ‫الحیویة‬ ‫الكیمیاء‬ ‫اختبارات‬ ‫عمل‬ ‫تم‬ ‫ذلك‬ ‫بعد‬ ‫‪.‬‬ ‫جنسن‬ ‫لوینستین‬ ‫وسط‬ ‫في‬ ‫زراعتھا‬ ‫والاستریتومایسین‬ ‫والایثامبیتول‬ ‫والایزونیازید‬ ‫الریفامیسین‬ ‫لأدویة‬ ‫الحساسیة‬ ‫تجارب‬ ‫اجراء‬ ‫تم‬ ‫كما‬ ‫‪.‬‬ ‫علیھا‬ ‫‪.‬‬ ‫العلاج‬ ‫في‬ ‫عملة‬ ‫المست‬ ‫للأدویة‬ ‫الجراثیم‬ ‫ھذه‬ ‫مقاومة‬ ‫لمعرفة‬ ‫في‬ ‫بمثیلاتھا‬ ‫ومقارنتھا‬ ‫المعزولة‬ ‫الأنماط‬ ‫تمییز‬ ‫بغرض‬ ‫ار‬ ‫تي‬ ‫ان‬ ‫في‬ ‫وال‬ ‫الاسبولیقوتایبینق‬ ‫اجراء‬ ‫تم‬ ‫‪.‬‬ ‫الجرثومة‬ ‫وأصل‬ ‫منشأ‬ ‫وتحدید‬ ‫العالم‬ ‫أنحاء‬ ‫مختلف‬ ‫النتائج‬ ‫المقاومة‬ ‫نسبة‬ ‫كانت‬ ‫كما‬ ‫جنسن‬ ‫لوینستین‬ ‫وسط‬ ‫في‬ ‫الزراعة‬ ‫في‬ ‫موجبة‬ ‫نتائج‬ ‫أعطت‬ ‫العینات‬ ‫من‬ ‫‪58.7%‬‬ ‫‪.MDR-‬‬ ‫الأدویة‬ ‫من‬ ‫نوعین‬ ‫من‬ ‫لأكثر‬ ‫مقاومة‬ ‫المعزولة‬ ‫الجراثیم‬ ‫من‬ ‫‪%‬‬ ‫‪8.7‬‬ ‫أظھرت‬ ‫‪.%‬‬ ‫‪20‬‬ ‫دویة‬ ‫للا‬ ‫البیانات‬ ‫قاعدة‬ ‫فى‬ ‫جزیئى‬ ‫تصنیف‬ ‫دون‬ ‫منھا‬ ‫‪70‬‬ ‫تبقت‬ ‫فیما‬ ‫نوع‬ ‫‪28‬‬ ‫فى‬ ‫الجزیئى‬ ‫التصنیف‬ ‫نجح‬ ‫‪TB‬‬ ‫‪.‬‬ ‫الدولیة‬ ‫الاستنتاجات‬ ‫الزراعة‬ ‫عمل‬ ‫أھمیة‬ ‫رز‬ ‫تب‬ ‫وعلیھ‬ ‫السودانیین‬ ‫المرضى‬ ‫وسط‬ ‫عالیة‬ ‫للأدویة‬ ‫السلیة‬ ‫المتفطرة‬ ‫مقاومة‬ ‫نسبة‬ ‫‪.‬‬ ‫التشخیص‬ ‫في‬ ‫الجزیئیة‬ ‫الأحیاء‬ ‫وسائل‬ ‫ادخال‬ ‫أھمیة‬ ‫تبرز‬ ‫كما‬ ‫للأدویة‬ ‫الجرثومة‬ ‫مقاومة‬ ‫لتحدید