Objective To develop a method for rapidly identifying Clostridioides difficile (C. difficile) and determining high-producing toxin strains, conduct clinical evaluation. Methods The loop-mediated isothermal amplification (LAMP) method was used to identify C. difficile based on the tcdC, tcdA, and tcdB genes. The sensitivity, specificity, and overall consistency of the detection method were evaluated. Results Feces specimens from 499 hospitalized patients suspected of C. difficile-associated diarrhea were detected, with C.difficile detection rate of 12.8% (64/499), out of which the detection rate of toxin-producing C. difficile was 10.8% (54/499). The sensitivity, specificity, positive predictive value, and negative predictive value of the detection method for tcdA were 87.2%, 98.9%, 89.1%, and 98.6%, respectively, and 88.2%, 99.6%, 90.0%, and 98.73% for tcdB , respectively. The total toxin levels of different strains were different, but the average toxin production level of A+B+ strains (1.79 μg/mL) was higher than those of A-B+ strains (0.72 μg/mL) and A-B- strains (<0.10 μg/mL). Conclusion The portable high-throughput LAMP detection method can rapidly and efficiently identify C. difficile and determine high-producing toxin strains.