The  objective of this study was to evaluate isolation, and evaluate the polymerase chain reaction (PCR) technique to confirm diagnosis of Mycoplasma gallisepticum (MG) in broiler chickens. One of the finest independent organisms is MG, can be reproduced autonomously, the lack of a cell wall, allowed it to take on various shapes and sizes, and to resist cell-wall targeting antibiotics. When MG infect chickens it caused chronic respiratory disease (CRD), characterized by rales, sneezing, coughing, nasal discharges, dyspnea, conjunctivitis. Decreased feed intake, feed conversion, an increase in mortality, carcass damage and medication costs, causing high economic losses. Diagnosing the cause is the first step in treatment, for evaluation isolation and direct PCR a total of 180 tracheal swabs were collected from broiler chickens (28-40) days old who had symptoms of CRD, during the period (1/12/2022-28/2/2023). Prevalence of MG by, isolation and direct PCR was 30.5% (55/180) and 32.77% (57/180) respectively. The sensitivity and specificity of direct PCR were 100% and 96.8%  respectively. When comparing culturing with PCR , the study found that the sensitivity and specificity were 93% and 100% respectively. The study concluded that culturing is still the golden standard test for MG detection for its high sensitivity and specificity but takes a long time, direct PCR is very fast and efficient.