Sugarcane is the major source of sugar in Asia and Europe, grown primarily in the tropical and sub-tropical zones of the world. The main disease responsible for its low yield is red rot. Therefore, in the present study, characterization of red rot disease was performed among 55 different sugarcane cultivars varying in red rot resistance level. 18 fragments were found to be associated with red rot resistance and were identified as resistant specific markers. The resistant specific fragments were amplified by RGA169, RGA396, RGA129, RGA231, RGA251, RGA057, RGA118, RGA152, RGA327, RGA542, RGA012, RGA173, RGA184, RGA275, RGA019, RGA267, RGA281 and RGA533. 7 fragments were found to be associated with red rot susceptibility and were considered as susceptible specific markers amplified by RGA088, RGA162, RGA396, RGA231, RGA251, RGA087 and RGA275. Sequencing of five resistant fragments, viz., RGA169, RGA231, RGA251, RGA267 and RGA533 was performed and the data thus obtained showed 80-99% similarity when compared with other resistant gene sequences previously submitted in NCBI database.