Detection of β-lactamases by Neutralizing Antibodies

Faculty Veterinary Medicine Year: 2014
Type of Publication: Theses Pages: 174
BibID 11821219
Keywords : Veterinary Bacteriology    
Resistance to p-lactam antibiotics is an increasing problem worldwide. The emergenceof multidrug resistant bacteria is mainly associated with P- lactamase production, sodevelopment of new diagnostic and therapeutic methods is crucial to keep pace withcontinually evolving pathogens. The current research aims to diagnose and treat the P-Jactam bacterial resistance by using anti p-lactamase serum. To achieve this goal thefollowing has been done:1- E. coli and K. pneumoniae isolates obtained from different sources were resistant to~-lactams in addition to other drugs by disk diffusion sensitivity test.2- The CMY p-lactamases from a blacMY E. coli isolate was purified by lysozymebased method.3- The CMY p-lactamase extracted from standard isolate in addition to standard CMYenzyme were subjected to sodium dodecycle sulfate gel electrophoresis (SDS-PAGE)for their purity assessment and determination of their molecular weights. The proteincontent of the purified enzyme was determined by spectrophotometer.4- Polyc1onal anti-CMY p-lactamase antibodies could be produced in immunizedrabbits.5- Anti- CMY p-lactamase antibodies had the ability to combine with the extracted P-lactamase through indirect ELISA technique and detect CMY and many AmpC P-lactamases in a number of E. coli and K. pneumoniae isolates.6- Exposure of 10 resistant E. coli and K. pneumoniae isolates to p-lactams and otherdrugs under the effect of anti p-lactamase sera through broth rnicrodilution MIC testand total colony count to determine antisera effect and comparing the MIC value andcolony count before and after addition of antisera. 1- There was an irreversible relationship between age of broilers and relative humidity, ammonia and carbon dioxide.2- Highest temperature was recorded in summer (34.4°C±0.41), spnng(27.3°C±O.35), autumn (26.3°C±O.36) and winter (22.7°C±O.82).3- Highest relative humidity was recorded in winter (81.5% ±22.6)followed by spring (75.2%±1.64), autumn (71.5%±2.27) and summer(7 1.4%± 1.84).-c-4- Highest ammonia concentration m (ppm) was recorded in winter(lO.7±1.65), spring (9.3±O.83), autumn (8.7±O.99) and summer(6.4±O.64).5- Highest carbon dioxide concentration in (celL) was reported in winter(3.78±0.42), followed by spring (2.92±O.22), summer (2.53±O.22) andautumn (2.23±O.3).11- Occurrence of airborne bacteria in broiler houses:1- There was a steady increase in the viable count of airborne bacteriathroughout the fattening period of broiler. The maximum bacterial 

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  • Eman Abdel Hameed Abdel Fattah Mohammed, "Detection of β-lactamases by Neutralizing Antibodies", 2014 More

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