University of Khartoum

Frequency of Fecal Carriage of Multidrug Resistant E.coli and Klebsiella Pneumoniae Among Hospitalized Patients and Healthy Volunteers

Frequency of Fecal Carriage of Multidrug Resistant E.coli and Klebsiella Pneumoniae Among Hospitalized Patients and Healthy Volunteers

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Title: Frequency of Fecal Carriage of Multidrug Resistant E.coli and Klebsiella Pneumoniae Among Hospitalized Patients and Healthy Volunteers
Author: Elhassan, Alaa Mohammed Mahmoud
Abstract: Background: Widespread use of antibiotics in Sudanese community and hospitals has led to emergence of high level of different types of antibiotic resistance among isolates of Enterobacteriaceae. Intestinal colonization with multidrug resistant Escherichia coli and Klebsiella pneumoniae is associated with high risk of developing infections such as urinary tract infection and wounds infection, so that the detection of fecal carriage of multidrug resistant Enterobacteriaceae is important in patients and in healthy people. The aim of this study was to determine fecal carriage of multi drug resistant (MDR) and extended spectrum beta lactamase (ESBL) producing E.coli and Klebsiella.pneumoniae among hospitalized patients in Soba University hospital and healthy volunteers from the community and define the nature of bla genes among them. Methods: A total of 64 hospitalized patients on antibiotics and 94 healthy volunteers who did not receive any antibiotics for 3 months were included in this study. Their age ranged from 10 to 60 years with a mean age of 35years. Male to female ratio was 1:1. Stools specimens were collected in sterile leak proof containers and transported to the lab within one hour.: Specimens were inoculated on MacConkey agar plates and incubated overnight at 37c°.Lactose fermenting colonies were isolated, Gram stained isolates were subjected to: indole test, citrate test, ureasa test, kligler iron agar and MR /VP test. K.pneumoniae was Gram negative bacilli, citrate +ve and VP +ve. E.coli was Gram negative bacilli, indol +ve and MR+ve .Seventy eight E.coli were isolated from the healthy volunteers, 44 E.coli and nine K.pneumoniae were isolated from the hospitalized patients. Antibiotics susceptibility test was performed by disc diffusion method, dissolving 1-2 colonies in peptone water and turbidity was compared with 0.5 MacFarland turbidity standard. Using sterile swab the suspension was inoculated onto Mueller- Hinton agar plate and antibiotics discs were placed (ciprofloxacin, gentamicin, tetracycline). Plates were incubated overnight at 37c°.Zone of inhibition was read and compared with VI comparable charts of control strains to determine the isolate as sensitive, resistant or intermediate sensitive. ESBL screening test by using cefpodoxime disc, when zone of inhibition ≤17mm this indicated the isolate was ESBL positive and confirmatory ESBL test should done. ESBL confirmatory test was done by double disc diffusion method using amoxicillin –clavulanic acid in the center and ceftazidime, cefotaxime away 20mm in each side, key hole between ceftazidime and \or cefotaxime with amoxicillin- clavulanic acid indicated ESBL production. DNA extraction was done by Gene MATRIX Bacterial & Yeast Genomic DNA Purification Kit, Characterization of ESBLs genes was performed by multiplex PCR using TEM,SHV,CTXM bla gene primers.. Result: Out of 78 E.coli isolate from healthy volunteers ESBL fecal carriage was found in 5(6.4%). TEM bla gene was detected in 5(100%) CTX-M bla gene 4(80%); SHV bla gene was not detected . ESBL-producing isolates were resistant to ciprofloxacin (10%) and tetracycline (4.4% ) MDR was observed in 9 (11.2%). Out of 53 isolate from hospitalized patients ESBL fecal carriage was found in 33 (62.4%). TEM bla gene was found in 25 (100%) CTX-M bla gene 17 (68%). SHV bla gene 6 (24%). ESBL-producing isolates were resistant to ciprofloxacin (83.3%), tetracycline (70%) and gentamicin(83.3%) .MDR was observed in 18 ( 33.9%). Conclusion: The prevalence of fecal carriage of MDR and ESBL -producing E. coli and K.pneumoniae detected in this study is much higher in hospitalized patients than healthy volunteers. Third generation cephalosporines in hospitals is considered as the most cause of gut colonization. Antibiotics misuse and poor hygiene are responsible for spread of MDR and ESBL producing Entreobacteriacaea in communities. Resistant to other classes of antibiotics is more observed in ESBL producer. TEM bla gene is the most spreading gene in fecal flora
URI: http://khartoumspace.uofk.edu/123456789/25586


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