Starter and non starter lactic acid bacteria (NSLAB) are responsible for enhancing unique flavour properties in traditional dairy products such as Lighvan cheese through biochemical mechanisms. In addition to saving domestic genetic resources, industrial production of semi-traditional dairy products is one of the major objectives of non starter microflora identification in indigenous dairy products. Previous studies based on the conventional methods performed in this laboratory showed that lactobacilli were the main microbial group colonizing in this type of cheese after 4 months ripening. In this study, eight batches of Lighvan cheese at 4 months ripening period were purchased from the local shop in the Lighvan village, Tabriz province. At the microbial level the sorbitol agar was used to detect non starter facultatively heterofermentative lactobacilli. The results of chemical evaluation of cheese including moisture and NaCl content and acidity and pH value showed that Lighvan is belonged to the group of semi hard cheeses. At the microbial level the group of Lactobacillus plantarum was identified but it was impossible to distinguish the particular species of this group present in the cheese. It was decided to sort the problem out using rec A gene sequence. Amplification products obtained on agarose gel created three distinct band with 318 bp size for Lactobacillus plantarum , 218 bp size for Lactobacillus pentosus and 107 bp size for Lactobacillus paraplantarum . Results obtained using rec A gene revealed that 86% of Lactobacilli isolates classified as Lb. pentosus and 14% as Lb. plantarum .