Abstract : Klebsiella pneumoniae bacteria were isolated from different sources; urine, blood, burns and wound swab, sputum, ear swabs, fluid, pus, endotracheal swabs, CSF and abscess of biopsy as (37, 24, 16, 9, 4, 3, 3, 2, and 1) % respectively. It showed different levels of antibiotic resistance with highest level of resistance against Ampicillin (100 %), also it showed that 53.13% of isolates were positive and 46.87% were negative for producing of β-lactamase. Biosynthesized Ag-NPs has inhibitory action (MIC) against K. pneumoniae with concentration 10 mM and sub- MIC with concentration 5mM. XRD spectrum of silver nanoparticles exhibited at 2ø (32.5°) corresponds to the (101) plane of conventional XRD data of Ag- NPs Crystals, while FTIR analysis was showed that stretch for Ag-NPs discovered around 518.58 cm-1. All isolates (100%) were positive to htrA, entB, blaCTX, and blaTEM before treatment with silver nanoparticles, but genes of htrA, entB, and blaTEM disappeared in the isolate no.1 after treatment with Ag-NPs at concentration (5mM). After gene sequence, the results have shown that before treatment with AgNPs, htrA, entB, blaCTX, and blaTEM genes were identical (100%) with NCBI, while after treatment with AgNPs in concentration (5 and 10) mM these genes were not affected.