Chang_2026_Sci.Rep__

The relationship between phenotypic resistance to organophosphate and pyrethroid insecticides and the frequencies of knockdown resistance (kdr) and acetylcholinesterase (ace-1) target-site mutations was investigated in four field strains of Anopheles sinensis collected from high-malaria-risk areas in the Republic of Korea (ROK). Six kdr genotypes and six ace-1 genotypes were identified, and the overall frequencies of resistance-associated mutations ranged from 62.1% to 96.8% for kdr and from 84.5% to 100% for ace-1. Among larvae that survived exposure to LC(50) concentrations of selected insecticides, resistance-associated genotypes, including Phe-Phe (TTT) in kdr and Gly (GGR)-Ser (YGC) in ace-1, were predominantly detected. Although the mean frequency of ace-1 resistance-associated mutations was higher than that of kdr mutations, the difference was not statistically significant (P = 0.42). In contrast, bioassay results showed that the mean resistance levels to pyrethroids were significantly higher than those to organophosphates (P = 0.006). Correlation analyses revealed no statistically significant association between kdr mutation frequency and pyrethroid resistance or between ace-1 mutation frequency and organophosphate resistance. These findings suggest that resistance-associated target-site mutations are widespread in An. sinensis populations from high-malaria-risk areas near the DMZ, but their frequencies alone do not fully explain phenotypic resistance levels. Therefore, while molecular data are useful for monitoring the distribution of resistance-associated mutations, they should be interpreted alongside bioassay results when developing insecticide resistance management strategies.