D3226: Comparison of three PCR-based methods for detection of SNP associated with pesticide-resistance

Recently, molecular diagnostic methods have been developed to detect frequency of SNP associated with pesticide resistance in a population. In this study, we developed two methods using quantitative sequencing (QS) and digital droplet PCR (ddPCR) to investigate G119S mutation in Haemaphysalis longicornis acetylcholinesterase1 (HlAChE1), and compared the detection efficiency between two methods using plasmid containing G119S mutation. According to QS, the peaks of chromatogram showed significant correlation with actual mixture of different ratio of G119 and S119 as judged by linear regression. The minimum detectable frequency of G119S mutation was 10% in QS. For ddPCR, probes for detection of G119 and S119 sites were labeled with FAM and HEX fluorescence, respectively, and at least 1% of mutation was detectable by ddPCR. These two methods developed in this study can be applied for molecular diagnosis of G119S mutation in HlAChE1.