Background Syphilis remains a significant public health concern in India. Ensuring the accuracy of diagnostic tests is crucial for effectively managing this disease. Objectives This study aims to assess the detectability of syphilis using commercially available non-treponemal and treponemal tests due to observed discrepancies in test results, which can lead to confusion and anxiety among healthcare providers and patients. Materials and methods We analyzed 2312 serum samples using the rapid plasma reagin (RPR), Treponema pallidum hemagglutination assay (TPHA), enzyme-linked immunosorbent assay (ELISA), and modified TPHA rapid test, interpreting the results according to the manufacturers' instructions. We evaluated the diagnostic accuracy of all four tests. Concordance between the traditional and reverse algorithms was determined by calculating the percentage of agreement and the kappa (κ) coefficient. Results Of the 2312 samples tested, 34 (1.5%) were positive, and 2098 (90.7%) were negative across all four tests. Comparing the test results with clinical diagnosis, TPHA and TP-ELISA showed the highest sensitivity at 96.08%, while RPR demonstrated the highest specificity at 100%. The agreement between the traditional and reverse algorithms was moderate, with a 97.3% agreement and a κ value of 0.53. Conclusion Reliance on a single serological test for syphilis screening presents limitations. A combined approach using both RPR and TPHA tests can more accurately diagnose and confirm syphilis. This combination strategy is cost-effective and relatively simple to implement.