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Molecular Detection of CTX-M Genes in Klebsiella pneumoniae Isolated from Different Clinical Samples in Baghdad City

Author: Ahmed Salim Mohammed
Journal: Medical Journal of Babylon مجلة بابل الطبية ISSN: 1812156X 23126760 Year: 2015 Volume: 12 Issue: 1 Pages: 152-160
Publisher: Babylon University جامعة بابل

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Abstract

CTX-M extended-spectrum β-lactamase (ESBL) producing Klebsiella pneumoniae have been reported to be an important nosocomial infections. A total of 50 K. pneumonia isolates were isolated from different clinical samples in some public hospitals in Baghdad city during the period from October to December 2013. Bacterial identification was done using conventional cultural & chemical methods &and VITEK 2 cards (GN) for identification, while the antimicrobial drug susceptibility of K. pneumoniae was performed by disk diffusion test and the minimum inhibitory concentration (MIC) testing was performed using VITEK 2 automated system (bioMérieux, France). ESBL production was phenotypically detected by double disk synergy test according to the Clinical and Laboratory Standards Institute(CLSI) guidelines. The presence of bla-gene encoded CTX-M was detected by conventional PCR technique.Out of 50 K. Pneumonia isolates,13 (26%) were ESBL producer by CDT, the minimum inhibatory concentration (MIC) of different antibiotics was performed on these 13(26%) isolates using VITEK2 AST-GN30 showed that 13 (100%) isolates were Ceftazidime,Ceftraiaxone and Cefepime resistant with MIC ≥16- ≥64 µg/ml,and 8(61.53%) of ESBL producing isolates were carbapenem sensitive 8 (61.53%) with MIC <=0.25 µg/ml. PCR assay revealed that 4 (30.76%) of the ESBL producing isolates harbored blaCTX-M gene.Extended spectrum beta lactamase mediated resistance in K. pneumonia is a cause for concern in the therapy of critically ill patients. The ESBL producing K. pneumoniae isolates were more resistant to various antimicrobial agents. This suggests that ESBL producing isolates in hospitals may cause serious infections that illustrated when these strains were responsible for a nosocomial outbreak.The findings strongly suggest that there is a need to track the detection of ESBL producers and that judicious use of carbapenems is necessary to prevent the further spread of these organisms. The prevalence of multi-drug resistant K. pneumoniae isolates especially ESBL producing bacteria was increased in Baghdad city .Phenotypic and molecular characterization of ESBL provide information about the prevalence of ESBL producing K. pneumoniae in Baghdad. The blaCTX-M was one of the predominant ESBL genes in K. pneumoniae in this study.


Article
Prevalence of Metallo-β-Lactamaseproducing Pseudomonas aeruginosaisolated from different clinical samples in Baghdad province

Authors: Alaa H. Al-Charrakh علاء الجراخ --- Salwa Jaber Al-Awadi سلوى جابر العوادي --- Ahmed Salim Mohammed احمد سالم محمد
Journal: Al-Qadisiyah Medical Journal مجلة القادسية الطبية ISSN: 18170153 Year: 2014 Volume: 10 Issue: 18 Pages: 13-19
Publisher: Al-Qadisiyah University جامعة القادسية

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Abstract

Background: Metallo beta lactamase(MBL) producing Pseudomonas aeruginosahave been reported to be an important nosocomialinfections.Pseudomonas aeruginosa is a leading cause of nosocomial infections, giving risetoa wide range of life-threatening conditions. Its intrinsic & acquired resistance to many antimicrobial agents and its ability to developmultidrug resistance imposes a serious therapeutic problem.Materials and Methods: A total of 75 P.aeruginosaisolates were isolated from different clinical samples in some public & private hospitals in Baghdad city during the period from April to August 2011.Bacterial identification was done using conventional cultural & chemical methods &and VITEk 2 cards for identification (GN), while the minimum inhibitory concentration (MIC) testing was performed using disk diffusion,E-test for Imipenem& Meropenem(oxoid, UK) & (AST-GN30)cards in VITEK 2 automated system(bioMérieux, , France).Each P.aeruginosa isolates showed resistance to Carbapenems(Imipenem& Meropenem) were subjected to Imipenem-EDTA combined disc synergy test (CDST) to investigate the production of MBL(confirmative test)Results:Out of 75 P.aeruginosa isolates,16 (21.3%) were grow on MacConkey agar supplemented with Meropenem4mg/L (MMAC),this method used as screening test, The MIC of different antibiotics was performed on these isolates using different methods(VITEK2AST-GN30,Imipenem&Meropenem E-test) showed that 6 (37.5%) isolates were Carbapenem resistant MIC ≥16µg/ml,while 4(25%) pseudomonas isolates appear to be MBL producer usingImipenem-EDTA combined disc synergy test(CDST). Discussion: MBL mediated carbapenemresistance in P. aeruginosa is a cause for concern in thetherapy of critically ill patients. The MBL producing P. aeruginosa isolates were more resistant to various antimicrobial agents. This result suggests that MBL producing isolates in hospitals may cause serious infections that illustrated when these strains were responsible for a nosocomial outbreak.The findings strongly suggest that there is a need to track the detection of MBL producers and that judicious use of carbapenems is necessary to prevent the further spread of these organisms.Conclusion: The prevalence of multi-drug resistant P.aeruginosa isolates especially Carbapenem resistant bacteria was increased in Baghdad province.Phenotypic characterization of MBLs provide information about the prevalence of MBLs producing P. aeruginosa in Baghdad.

بكتريا الزوائف الزنجارية المنتجة لانزيم Metallo-β-lactamase مثبته بانها من المسببات المهمة للاصابات والالتهابات المختلفة في المستشفيات. بكتريا الزوائف الزنجارية لها قابلية عالية على مقاومة العديد من المضادات الحياتية المستخدمة حاليا من خلال ميكانيات مختلفة منها داخلية او مكتسبة واهم الميكانيكيات هو انتاج انزيم β-lactamaseMetallo- تم جمع 75 عزلة من بكتريا الزوائف الزنجارية من مختلف العينات السريرية من بعض المستشفيات والمختبرات الحكومية والخاصة في محافظة بغداد للفترة من نيسان ولغاية اب لعام 2011. تم تشخيص البكتريا باستخدام مختلف الطرق سواء كانت فحوص كيميائية او عن طريق تشخيص البكتريا في اوساط زرعية مختلفة,كذلك تم تشخيص العزلات البكتيرية باستخدام جهاز VITEK2 وتم قياس الحساسية الدوائية للعزلات البكتيرية باستخدام disk diffusion وكذلك تم قياس اقل تركيز مثبط من العقار (MIC) باستخدام جهاز VITEK2 وكذلك طريقة E-Test لعقاريImipenem وMeropenem وتم اجراء فحص (CDST) كفحص تاكيدي للعزلات المقاومة لعقاري Imipenem ,Meropenem للتاكد من انتاج انزيم MBL .وتم التوصل للنتائج التالية:نسبة انتشار العزلات المقاومة لعقارات الكاربابينيم هي 8% وعدد العزلات البكتيرية المنتجة لانزيم MBL باستخدام الطرق المظهرية هو 4 وبنسبة 5.3%.

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