Dahi is a fermented milk product containing probiotic lactic acid bacteria. This study aimed to isolate, identify, and characterize lactic acid-producing bacteria from native Dahi and evaluate their viability in orange juice using natural prebiotics. Dahi samples were obtained from local shops in Chattogram and Bogura, Bangladesh. Lactic acid-producing bacteria were isolated using MRS (de Mann Rogosa and Sharpe) medium. The isolated bacteria were identified through colony morphology, biochemical tests, and probiotic characteristics. Molecular identification was performed using polymerase chain reaction (PCR) targeting conserved 16S rDNA regions. Isolates of the genus Lactobacillus and Lactococcus lactis sp. Lactis were confirmed and used to develop probiotic orange juice. Prebiotics (Asparagus falcatus and Zingiber officinale) were added to the juice to support probiotic growth. The inoculated cell’s viability and the juice’s physicochemical parameters were evaluated during fermentation (48 hours) and storage (28 days). All fruit juice samples showed a mean number of viable cells of at least 1×105 CFU/mL during the 48-hour fermentation and 28-day storage in the refrigerator. Using natural prebiotics positively affected the survival of lactic acid bacteria, as demonstrated by bacterial colony growth on Petri dishes. Developing probiotic fruit juice enriched with prebiotics could be an effective alternative for individuals allergic or intolerant to milk-based products. Incorporating lactic acid bacteria from native Dahi into orange juice, combined with natural prebiotics, resulted in viable probiotic cells throughout fermentation and storage.