Human leukocyte antigens (HLA) are genetically derived proteins in the major histocompatibility complex. They help distinguish “self” and “non-self” antigens and are essential in interacting with the immune cells inside the body. The present research work examines the prevalence of HLA-B27 among patients suspected of Spondyloarthritis (SpA), which has also been correlated with Interleukin-17/23 Inflammatory Markers and other clinical manifestations and was carried out between August 2017 to January 2021. The patient’s blood samples were collected and tested for HLA-B27 and Interleukin-17/23 inflammatory markers. Among 289 SpA patients, 60% (172) were males, and 40% (117) were females, with a ratio of 1.5:1. Ankylosing Spondylitis (65.1%) was found to be the most prevalent subgroup of SpA among the patients, closely followed by reactive arthritis (21%), psoriatic arthritis (10.7%), undifferentiated spondyloarthritis (2.1%), and inflammatory bowel disease with associated arthritis (1%). HLA-B27 was found to be positive in 54% (156) out of 289 patients. Normal IL-17 ranges were seen in 42% of HLA-B27- positive patients, while increased IL-17 was seen in 58% of the population with positive HLA-B27 cases. IL-23 was found within normal ranges in 40% of positive HLA-B27 cases, while it was found to be increased in 60% of the positive HLA-B27 positive subjects. We concluded that HLA-B27 was found to be positive among more than half of the patient population with SpA. The early detection of HLA-B27 may aid in changing lifestyle to prevent Spondyloarthritides.