Among a food-associated pathogen, Listeria monocytogenes has become the most important one. It can cause a "high fatality rates 20–30%" in comprasion to another foodborne bacteria which causing dangerous disease called listeriosis. Listeria outbreaks are often linked to dairy products and milk. This article intends to review phenotypic and genotypic characters of L. monocytogenes in Assiut city. How common L. monocytogenes is determined in different sources of raw milk (20 market milk, 40 buffleo milk and 40 cow milk).  By detection of 3 virulence genes and its sequencing (hlyA, inIB and (prfA) can deterimined the pathogenic potential of the isolates. Firstly enrichment samples in fraser broth, then, plating on to ALOA®, Merck. Finally a multiplex-PCR,was used for identification of suspected colonies. The results revealed that 8 (40%), 6 (15%) and 11 (27%) of samples that collected from market, buffalo and cow raw milk were positive respectively. Antibacteerial sensitive test showing in vitro thai Listeria mmonocytogenes had a higher sensitivity to Sulfa methoxazoletrimthoprim (SXT) and Ciprofloxacin, (CIP) followed by Chloramphenicol (C), low sensitive to  Gentamicin (CN), it resisted Erythrommycin (E) and Amoxicillin (AX). The PCR results for isolates have hly A Inl B and prf A genes of L. monoctogenes.