Clostridium perfringens is a Gram positive anaerobe which makes spores that help it to persist under different environmental conditions. Spores of C. perfringens have the potential to remain in the fecal matter and soil for many years due to which control of transmission of pathogen becomes difficult. The pathogen is responsible for various diseases in both humans and animals. We determined the prevalence of C. perfringens in the soil of selected districts of Punjab province of Pakistan along with potential association of different categorical variables. A total of 970 samples were processed for real time PCR based screening of genome corresponding to cpa gene (alpha toxin gene) of C. perfringens. Only n=96 samples were found positive [9.89% with CI: 95% (8.018- 11.78), p-value< 0.05]. Highest prevalence was found in district “Chakwal” followed by district “Attock” and “Lahore”. The villages found positive for C. perfringens were evaluated for different risk factors including distance from the main road, animal market and water source; animal density, number of houses in the village and animal and human interaction at the sampling site. Analysis of the data revealed that animal and human interaction was the associated risk factor in most of the districts followed by distance from the water source, animal density and human population of the village. The study concludes that C. perfringens is prevalent in the soil of Punjab province and the prevalence varies with several risk factors in different districts. Further studies are required to find out most prevalent type of C. perfringens as well as effect of different categorical variables on prevalence of C. perfringens in other parts of the country.