For generations, individuals have been incorporating wild edible fungi into their diets. Precisely identifying mushrooms is essential for harnessing their potential benefits for human use. Specifically, the taxonomical identification of <i>Pleurotus</i> species is known for its complexity. In modern times, it is always necessary to use molecular analysis as a valuable approach for gaining a deeper understanding of the systematics and the delineation of species. Thus, this study confirmed the identity of two wild isolates of <i>Pleurotus djamor</i> (pink oyster mushroom) using the internal transcribed spacer sequences. The amplicons underwent sequencing, and the resulting data was assembled and analyzed. The nucleotide sequences of the two wild isolates blasted against sequences from GenBank database revealed that two wild isolates matched 96.80 and 95.27% <i>P. djamor</i> with accession number KF280324.1 and KT273366.1. Commercially cultivated species took less time for spawn run (17.75 days), primordial initiation (22.25 days) and recoded highest biological efficiency (73.92%) among wild isolates. Nutritional analysis revealed that wild isolates showcase an abundance of nutrients, surpassing commercially cultivated species in terms of moisture, protein, and crude fiber content. This distinctive nutritional profile positions wild isolates as an exceptional superfood, fostering overall health enhancement.