Background: Malocclusion shares the third highest prevalence among oral pathologies, second to tooth decay and periodontal diseases. Objectives: In general, many studies have been conducted to find out the association between blood groups and oral and dental pathologies, for instance, some have succeeded; others have not due to the geographic diversity. So the study aimed to explore the relationship of ABO blood groups with classes of malocclusion and relative prevalence of these characters in males and females of a defined population. Method: The current study was performed on 500 individuals with an age range of 15 to 45 years, along with permanent dentition only, reporting to the department of orthodontics of Punjab dental hospital, Lahore. Complete details about the classes of malocclusion and blood group of the subjects were noted in accordance with the ABO blood group system. The sample was divided into three groups, i.e., Class I, Class II, and Class III. Data were analyzed statistically by using SPSS 21.0. Pearson Chi-square was used as statistical analysis to get the association of classes of malocclusion with blood group type. Result: A significant difference between the prevalence of classes of dental malocclusion in relation to blood group types was found among both genders. Conclusion: In conclusion, there was gender wise association difference between the classes of dental malocclusion and blood group types. The study also covered the gender wise prevalence of dental malocclusion in all the four blood groups which are A, B, AB and O. The new findings will be helpful for clinicians with regards to preventive orthodontics. The research findings are a good and informative addition to the present literature as well as provides a ground for further research studies in the same area.