The demand for fresh produce is rising daily as it's an excellent way to maintain a healthy lifestyle. The current work fixed the target employing culture-based and molecular techniques for isolating and identifying the pathogenic Vibrio parahaemolyticus from fresh produce. The colonial appearance on TCBS agar and CHROMagar culture plates revealed the prevalence of Vibrio spp. and V. parahaemolyticus in all examined samples, which were 45/112 (40.2%) and 40/112 (35.7%), with the highest load of up to 2.0x105cfu/g and 6.4x103cfu/g, respectively. Sixty isolates in all were chosen for further molecular characterization. Multiplex PCR results exhibited that all tested isolates were positive for tlh gene and six for tdh gene but none of isolate for trh gene. According to obtained data on antimicrobial susceptibility, the major portion of tested strains was found to be resistant to ampicillin (90%), tetracycline (80%), and streptomycin (70%). The range of the multiple antibiotic index was 0-0.33. This study finding revealed that the existence of potential pathogenic V. parahaemolyticus in tested fresh produce samples constitutes a risk to public health and consumer safety thus urging continued surveillance. [ J Adv Biotechnol Exp Ther 2024; 7(2.000): 303-313]