The aim of this study was to carry out a serological and molecular survey for the presence of Dirofilaria immitis infection in stray cats using an ELISA kit and PCR assay. One hundred and fifty-five stray cats (77 females and 78 males) in Gyunggi province in South Korea, were used in this study. Four (2.6%) tested with the ELISA kit showed a positive reaction, and all positive samples by the ELISA kit showed a positive reaction by PCR analysis. No significant difference was observed between the male (2.6%) and female (2.6%) cat groups by ELISA kit. The positive rates for dirofilariosis were 2.8% in the 4–6-year-old group, and 18.2% in the >6-year-old group by ELISA kit. With regard to the age element, older cats showed a higher prevalence of D. immitis infection in this study. A statistical analysis revealed that significant difference was observed in >6-year-old group (p<0.01). In conclusion, D. immitis infection in stray cats was present in Gyunggi province, although its incidence was low. Therefore, heartworm treatment and/or prophylaxis for stray cats captured are required in this area.