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Due to the development of acaricide resistance in Tetranychus urticae, a rapid and accurate monitoring method for acaricide resistance gene frequency is required for resistance management.In this study, we developed a new diagnostic gene frequency prediction method, using quantitative real-time PCR with allele-specific primer sets, for mutation of I1017F in chitin synthase I, G126S in mitochondrial cytochrome b, and H110R in the PSST subunit of mitochondrial electron transport complex I, which are involved in etoxazole, bifenazate, and pyridaben resistance in T. urticae, respectively. Resistance allele frequencies computed using the 2 −ΔΔCq method, in mixtures of resistance and wild-type (susceptible) alleles, were strongly correlated with the actual frequencies of the resistance alleles in DNA samples for all three acaricides (regression slopes of 0.945-0.996; R 2 > 0.99). This result strongly indicates that resistance allele frequency in a population can be accurately predicted using a diagnostic quantitative real-time PCR method with a resistance allele-specific primer set. Finally, we applied this method to nine field populations and successfully determined resistance allele frequencies for the three acaricides. Overall, this diagnostic prediction method may contribute to the development of acaricide resistance management strategies, via monitoring resistance allele frequencies for a wide range of acaricides.

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