الفهرس 
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Abstract
Escherichia coli and K.pneumoniae represents, one of the important nosocomial infections, have become a matter of major concern in MUHs. (12.6% 17.1%) of total pathogens respectively. The aim of this study was to detect the frequency of E.coli and K.pneumoniae causing nosocomial infections in MUHs. with detection of different factors increase the risk of their acquisition. Also detection of strains that produce ESβLase enzyme as a major cause of treatment failure in hospital infected patients and study phenotypic and genotypic characterization of such strains. Phenotypic characterization was performed using cultural characteristics and morphological appearance in Gram stained smears and conventional biochemical reactions. API 20E was used to confirm isolation to detect different species of E. coli and K. pneumoniae. Escherichia coli and K.pneumoniae were isolated especially from patients with abuse of broad spectrum antibiotics, immunecompromized conditions e.g. chronic renal failure, malignancy, diabetes, radiotherapy, steroid and in those with various types of instrumentation. They were also at high incidence in old age and in neonates. Respiratory tract infections and urinary tract infections are associated with long term endotracheal intubations and indwelling catheters. Infections at such site may lead to septicaemia. Because of advances in supportive care and therapy, large number of hospitalized patients is susceptible group likely to develop nosocomial infections due to E.coli and K.pneumoniae. Resistogram using Kirbey Bauer method was done to study the antibiotic susceptibility of the isolates. All the isolated strains show sensitivity to imipenem and most of them were sensitive to amikacin, while the least effective antibiotics are ampicillin. Screening for ESβLs was performed using double disk synergy test, and modified three dimension test to their production were detected in 143 cases at a percentage of 35.8% in E coli and 215 cases at apercentage of 39.7% in K. pneumoniae. PCR of all ESBL positive isolates (n=50) of E.coli and K. pneumoniae. revealed that 867 bp of TEM and 867 bp of SHV for E. coli and 867 bp of TEM and 930 bp of SHV for K. pneumoniae. Phenotypic methods are only screening methods for detection of ESBL in a routine laboratory. The genotypic methods actually help to confirm the genes responsible for ESBL production. Phenotypic methods provide an efficient, rapid differentiation of ESBL in E.coli and K.pneumoniae and could be used as a rapid tool for epidemiological studies among ESBL isolates. It is of great importance that PCR reaction conditions, including DNA template concentration, annealing temperature and the choice of the primer pairs, should be standardized to avoid variations in PCR patterns. Tracing the sources and revising our MDICU policies were strictly implemented, and helped us to control an infection caused by ESBL- E.coli and K.pneumoniae and eliminate serious carbapenem resistant E.coli and K. pneumoniae strain. Escherichia coli and K.pneumoniae infections in MUHs. are caused by a variety of strain genotypes. It can help further efforts to understand the spreading of this bacterium.