This study evaluated the nature to bacterial infections in a neonatal intensive care unit (NICU) at Zagazig University Hospital, Zagazig , Egypt . Rapid diagnosis and identification of the infectious bacterial pathogens by polymerase chain reaction (PCR) is recommended herein. Out of 70 microbial cultures ordered by physicians from many clinical samples, 50 cultures showed bacterial growth. The 50 bacterial cultures obtained were identified by biochemical methods. They were categorized into 6 groups ; were arranged in the following descending order according to number the identified strains : Escherichia (E.) coli (group 1 , 22 isolates) > Staphylococcus (S.) aureus (group 2 , 10 isolates > Streptococcus (St. ) pneumonia (group 3 , 6 isolates) > St . agalactiae (group 4 , 5 isolates) > St. pyogenes (group 4 , 5 isolates) > Klebsiella (Kb.) pneumonia (group 6 ,3 isolates) . Antibiotic sensitivity profiles showed variability in susceptibility of isolates to the 10 studied antibiotics. Certain virulence genes for selected bacterial strains from each bacterial group were rapidly detected by multiplex PCR ; this indicated that PCR technique could be used as a rapid for identification of infectious bacteria and in turn a diagnostic tool for infectious diseases .It reduces time and personal efforts faced microbiologists throughout biochemical identification.

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