Molecular Characterization of Colistin Resistant Genes of Acinetobacter bumannii among ICU Patients in Khartoum State

Abstract

Abstract Background: A.bumannii is the most prevalent infection- in the hospital environment, which is considered problem in treatment, and the infections-caused by it are difficult to treat, because it is multidrug resistant. Aim: to detect colistin resistant genes ( MCR1, MCR2, MCR3,MCR4, MCR5) of A.bumannii isolated from different specimens from ICU in Khartoum state using Multiplex PCR Technique Methods: Clinical samples were collected, which yielded A.bumanii isolates from different hospitals, re-identified was done by phenotyping methods include Gram stain and biochemical and susceptibility tests DNA was extracted by boiling method and PCR was done using specific primers to detect the colistin resistance genes and visualized by U.V light. Result: In the present study out of the total number of pathogenic Gram-negative isolates were 650 of which 67 isolates were A. baumannii (10.3%),from different sample include sputum(30/67) 44.8% wound swab(9/67) 13.4%, blood (7/67) 10.5% urine (11/67)16.4% CVC tip (6/67)8.9%CSF(3/67)4.5%bedsore swab(1/67)1.5%.The most isolates from males (47/67)70.1% and (20/67) 29.9%from female, The isolates were highly resistant to tested antibiotics, cefepime, , ceftriaxone, cefuroxime, gentamicin, amikacin, ciprofloxacin, amoxiclav, aztreonam and and meropenem at 100 %, and resistance to colistin (42/67)(62.6%). PCR result showed resistant genes of colistin (MCR 1, MCR 2, MCR3, MCR 4 and MCR 5) were detected in (83.5%) (56/67) in percentage 76.7%(43/56), 44.6%(25/56), 3.6%(2/56), 1.7%(1/56) and 0.0%(0/56). Bioinformatics analysis showed that universal Genes of the seven sequences (sudan001, sudan002, sudan003, sudan005, sudan006 sudan007 and sudan008) were related closely to each other than sudan004. Also, these sequences shared a common ancestor were the Australia strain followed by China strain. Where Sudan 004 was related closely to the United Kingdom strain and the USA strain. Conclusions: Our present study concluded that the frequency of colistin resistance gene of A.bumanii was 83.5 %( 56/67) in different specimens collected from ICU patient in Khartoum state .

المستخلص ` خلفية الدراسة: تعد بكتيريا الراكده البومانيه أكثر الكائنات الحية المسببة للعدوى انتشارًا في بيئة المستشفى ، والتي تعتبر مشكلة خطيرة في علاج الالتهابات التي تسببها ، لأنها مقاومة للأدوية المتعددة. الهدف: الكشف عن الجينات المقاومة للكوليستين (MCR1، MCR2، MCR3، MCR4، MCR5) الراكده البومانيه المعزولة من عينات مختلفة من العناية المركزة في ولاية الخرطوم باستخدام تقنية Multiplex PCR الطريقة: تم جمع العينات السريرية ، والتي أسفرت عن عزلات الراكده البومانيه من مستشفيات مختلفة ، وتم التعرف عليها من خلال طرق التنميط الظاهري بما في ذلك صبغة جرام واختبارات الكيمياء الحيوية والحساسية واستخراج الحمض النووي بطريقة الغليان وتم إجراء تفاعل البوليميراز المتسلسل باستخدام بادئات محددة للكشف عن الجينات المقاومه للكوليستين. وتصور بواسطة ضوء الأشعة فوق البنفسجية.