The goal of the present study was to investigate the potency and validity of cold atmospheric pressure plasma (CAP) in combating bacterial contamination and bacterial biofilm for wound healing. CAP is applied as a topical treatment with the hope of providing a less invasive approach compared to surgical debridement and irrigation. The setup of helium CAP jet has been done with 3.4 KV and frequency 16.1 KHz. The antibacterial activity of the plasma jet was performed on Muller Hinton agar media (MHA) for Staphylococcus aureus, Bacillus cereus, and Escherichia coli, but selective Enterococcus agar media (Bile Esculin Agar Media) (BEA) was used for Enterococcus faecalis. CAP treatment (1, 2, and 3 min) showed that plasma jet can inhibit and limit the growth of pathogenic G +ve (Enterococcus faecalis, Staphylococcus aureus, and Bacillus cereus) as well as G –ve bacteria (Escherichia coli) depended directly on the strains and also depended greatly on the exposure. The plasma jet has antibacterial potency against Gram-Positive Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus, and Staphylococcus aureus in addition to Gram-Negative bacteria Escherichia coli.