الكشف عنcag جين وجينات الضراوة الاخرى من بكتريا القولون المسببة لالتهاب المجاري البولية في مدينة بابل == Screening for cag Genes and Other Virulence Genes Among Escherichia coli Isolated From Patients With Urinary Tract Infection In Babylon City
Author name:
ياسمين رياض سعيد العزاوي
Supervisor name:
محمد صبري عبد الرزاق السعيد | محمد رضا جودي
General topic:
Medicine
Specific topic:
Microbiology
Degree:
Master
University:
University of Babylon - Faculty Of Medicine
Language:
English
University location:
Babylon
First pages:
19T1672 - p.pdf
Abstract:
في هذه الدراسة، تم جمع (100) عينة ادرار من مرضى مصابين بالتهاب المجاري البولية و(25) عينة خروج من افراد اصحاء كعينة ضابطة، من كلا الجنسين في مستشفى الحلة الجراحي خلال الفترة من شباط 2015 الى ايار 2015.ثمانون عينة فقط اظهرت نمو على الاوساط الاختبارية. و40 عينة كانت حاملة ل E.coli بنسبة 10(25%) عزلة من الذكور و30(75%) عزلة من الاناث. اضافة الى ذلك تم تشخيص (7) عزلات E.coli من عينات الخروج لاشخاص اصحاء .اظهرت الدراسة ان 20(50%) من عزلاتUPEC تمتلك القابلية على انتاج الهيمولاسين خارخ الخلية على وسط اكار دم الانسان بنسبة 15(37,5%) الفا هيمولايسين و5 (12,5%) بيتا هيمولايسين . اما باقي العزلات 20 (50%) كانت غير قادرة ع انتاج الهيمولايسين.تضمنت الدراسة ايضا الكشف الجزيئي عن cag A جين لمشاهدة Sequence homology لهذا الجين في بكتريا H.pylori وE.coli بواسطة تقنية تفاعل البلمرة المتسلسل (PCR) حيث لوحظ ان كافة عزلات البكتريا اظهرت نتيجة سالبة لهذا الجين . وقد لوحظ ايضا نفس النتيجة ل cag E جين.اما التحري عن vir BII جين باستخدامprimer , فقد لوحظ ان 5(71,4%) عزلة بكتيرية من عينات الخروج و25(62,5 %) عزلة بكتيرية من عينات الادرار كانت ايجابية لهذا الجين , الذي لم يسبق دراسته في بكتريا E.coli . هذه النتيجة مؤشر لوجود sequence homology لهذا الجين مع جين بكتريا H.pylori .كما تم فحص عزلات UPEC )) للكشف عن وجود جينات الالتصاقpap, sfa, afaI) ) بواسطة (PCR). حيث وجد ان 26(65%) عزلة ادرار كانت تحملpap جين. و25(62%) عزلة تحمل sfaجين. بينما afaI جين يوجد في 17(42%).كذلك الكشف الجزئي عن جينات (cnf I and cnf II). ووجد ان cnf I كان ايجابيا في 21(52%) عزلة ، بينما cnf II لم يتم تحديده في اي عزلة.اظهرت الدراسة ايضا ان aer جين متواجد في 25(62,5%) من العزلات. اما hly جين فانه موجود في 18(25%) من العزلات.بالاضافة الى ذلك تم دراسة تاثير بعض المضادات الحياتية على عزلات بكتريا القولون المعوية. وقد وجد ان جميع عزلات هذه البكتريا حساسة بنسبة 100% الى Imipenem واقل حساسية 60% الى Nitrofurantoin و55% الى Amikacin . لكن بقية العزلات اظهرت نسب مختلفة من المقاومة لمضاد واحد او اكثر من المضادات الحيوية مثل Ceftazidim 80% وCeftriaxone 87,5 % , وبدرجة اقل من المقاومة لكل من Levofloxacin 62,5% وTrimethoprim - Sulfametoxazole 77,5% . | In this study, a total of (100) urine specimens obtained from patients suffering from Urinary Tract Infection and also, 25 stool samples were taken from healthy individual as control group were included, from both sexes who admitted to Al - Hilla Teaching Hospital Babylon during a period from February (2015) to May (2015). The urine and stool samples were then cultured on selective media to isolate Escherichia coli. Only 80 samples showed positive culture growth. It was found that only 40 isolate of E. coli were isolated where 10 (25%) of the isolates from male patient and 30 (75%) from female patients. Moroever, 7 isolates E.coli were isolated from stool samples of human individual.The ability of uropathogenic E.coi (UPEC) to produce extracellular hemolysin was studied, and it was found that 20 (50%) out of 40 isolates of E. coli were able to produce hemolysin on human blood agar, whereas15 (37.5%) were found α - hemolytic and 5 (12.5%) β - hemolytic. Other isolates of E. coli 20 (50%) had no ability to produce hemolysin.On the other hand, molecular detection of cytotoxic associated gene A (cagA) to show the sequence homology of this gene in Helicobacter pylori and E.coli. by PCR technique showed that all isolates of E.coli from urine and stool gave negative result for this marker, which confirm that there is no sequence homology of this gene with that present in H. pylori.The same results were seen with cytotoxic associated gene E (cagE) by using specific marker, where all isolates from urine and stool samples gave negative result to this gene. Moreover, the isolates of E. coli were subjected to show their possession of virB II gene, by using specific marker. It was observed that 25 (62.5%) of bacterial isolates from urine and 5 (71.4%) of bacterial isolates from stool gave the positive result for this gene, which is not previously detected in E. coli. This result indicate to found sequence homology of this gene with that present in H. pylori.Besides, the isolated UPEC were examind for the presence of the adhesion genes (pap, sfa, afaI) by PCR assays. It was found that pap gene was present in 26 (65%) isolates and sfa gene present in 25 (62.5%). While afaI gene present in 17 (42%) of isolates. Molecular detection of cytotoxic necrotizing factor I and II (cnf I and cnf II), was also studied by using specific markers. It was seen that cnf I gene was positive in 21 (52.5%) isolates of E. coli. While cnf II gene was not observed in any isolates of E.coli. Aerobactin (aer) and hemolysin (hly) genes, is also investigated by using specific markers; it was found that aer gene was present in 25 (62.5 %) of isolates and hly gene present only in 18(45%). Finally, the susceptibility of isolates to a variety of antibiotics has been investigated. It has been found that E. coli isolates were sensitive to imipenim at a rate (100%), and lesser to nitrofurantoin 60% and amikacin 55%. But most of them showed different mode of resistance to antibiotics used in this study as Trimethoprim - Sulfamethoxazole 77.5%, Ceftazidim 80%, levofloxacin 62.5%, Ceftriaxone 87.5%.
