Introduction: Metabolic syndrome (MetSyn) as defined by the latest Harmonised definition and the agreement between the Harmonised definition and other definitions is poorly studied among Malaysians. This study was conducted to determine and compare the prevalence of MetSyn according to the Harmonised, International Diabetes Federation (IDF) and National Cholesterol Education Program (NCEP ATPIII) definitions among Malay staff of Universiti Putra Malaysia (UPM). Methods: Subjects aged between 20 to 65 years were recruited by convenient sampling. Waist circumference, blood pressure, lipid profiles and fasting plasma glucose levels were assessed. The agreement between the Harmonised and other definitions was determined by Kappa statistics. Results: A total of 227 subjects with a mean ± SD age of 37.9±9.6 years participated in the study. The overall prevalence of MetSyn was 38.3%, 38.8% and 33.5% according to Harmonised, IDF and NCEP ATP III definitions, respectively. Generally, men had higher prevalence of MetSyn than women. The prevalence increased with age in both genders with a more progressive trend in women. Men in the age group of 20-39 years had a high prevalence of metabolic syndrome. A strong agreement was found between the Harmonised and the IDF definitions (Kappa index=0.991), and between the Harmonised and the NCEP ATP III definitions (Kappa index=0.857). Conclusion: Regardless of definitions used, the prevalence of metabolic syndrome in the study, especially in young men, was high and warrants further investigation. The Harmonised definition is suitable for diagnosing metabolic syndrome in any population with similar sociodemographic characteristics.