Human toxocariasis is a highly prevalent zoonosis worldwide but is underreported in most countries. This study was conducted to evaluate Toxocara canis seropositivity in different exposure sub-groups located in the Mardan, Swabi, and Nowshera districts of the Khyber Pakhtunkhwa province of Northwest Pakistan. A total of 400 blood samples were collected from males 15 years of age and older with no animals, with livestock, with dogs and/or cats living in the house, and from butchers and veterinarians or para-veterinarians. Serum was tested using a commercial ELISA kit for detection of IgG antibodies against T. canis. Proportion seropositive was presented for each group and differences between groups were evaluated using the chi-square or Fisher's exact test as appropriate. Possible risk factors obtained through administration of a questionnaire were also evaluated for each sub-population. Overall T. canis seroprevalence was 14.2%, with a significant difference found between the seroprevalence of individuals with no animals (5.0%; 5/100), individuals with dogs and/or cats living in the household (8.0%; 8/100), individuals with livestock (18.0%; 18/100), veterinarians or para-veterinarians (24.0%; 12/50), and butchers (28.0%; 14/50) (p < 0.001). Significant differences in seropositivity by income bracket, education level, and working in the fields were found for some sub-groups. Study findings demonstrate that certain sub-populations, in Northwest Pakistan, may be at greater risk of T. canis infection. Development and implementation of targeted preventive strategies may, therefore, be needed.