Necrotic enteritis caused by Clostridium perfringens has been linked to severe economic losses in the poultry industry and represents a serious threat to public health due to spore-forming ability and its role as foodborne pathogen. Although chemical disinfection plays a role in preventing and controlling the spread of bacterial diseases in livestock and poultry housing, little is known about its efficiency against spore-forming C. perfringens. This study was designed to investigate the sporicidal activity of a combination of quaternary ammonium and glutaraldehyde disinfectant (TH4) against previously isolated ten C. perfringens strains according to European Standard EN 1276, 2019 application method at different concentrations (0.25%, 0.5%, 1%) with different contact times (0, 1, 5, 15, 30, 60 minutes). C. perfringens strains were identified, toxinotyped, and screened for the presence of QAC resistance genes (qacED1 and qacA/B). Five C. perfringens isolates were verified to produce toxinotype A, while the remaining five isolates were confirmed to produce toxinotype G. Screening for qacED1 and qacA/B resistance genes revealed their presence in six and four isolates, respectively. TH4 0.25% and 0.5% tested in this study didn't give the required five log10 CFU reduction in the spore count in five C. perfringens at all exposure times, while concentration of 1% achieved the required five log reduction in the count of C. perfringens spores for relatively long contact times of 60 minutes in five C. perfringens type A isolates. These findings recommend the use of concentration of TH4 1% and increase exposure time to 60 minutes to obtain effective sporicidal activity against C. perfringens strains.