In this study, the newly designed pqq gene-specific primer sets were used for determination of phosphate-solubilizing capabilities of bacterial isolates from the agricultural regions of Erzurum. The specificity of newly designed primer sets (PqqA2F/PqqA2R, Pqq5F/Pqq5R, PqqF2/PqqF2R) were tested against ten isolates, whose phosphate-solubilizing activities were initially proved by the conventional methods. Non-phosphate-solubilizing bacteria were also chosen as negative control. According to the results, five of ten phosphate-solubilizing bacteria with PqqA2F/PqqA2R, two of ten phosphate-solubilizing bacteria with Pqq5F/Pqq5R primer set, and one of ten phosphate solubilizing with PqqF2F/PqqF2R bacteria were successfully amplificated in the PCR assay and none of the non-phosphate-solubilizing bacteria was amplificated. Then, the molecular characterization of the active phosphate-solubilizing strains was done based on the partial 16S ribosomal RNA gene region sequence analysis method. Two isolates of Enterobacter sp., 1 Rhizobium sp., 1 Enterococcus sp., 1 Bacillus cereus, 1 Bacillus atrophaeus, 1 Bacillus aryabhattai, 1 Acinetobacter sp., 1 Pseudomonas japonica, and 1 Enterobacter cloacae were identified as active phosphate-solubilizing strains. Consequently, the results showed that this specific primer sets could be used as an economic, rapid, and useful tool for the detection of phosphate-solubilizing strains in the agricultural researches.