关键词: 高分辨率熔解曲线, 家蝇, 抗性, 基因突变, cyp6d1

Abstract: Objective To investigate the mechanism of deltamethrin resistance in Musca domestica. Methods The mutation and amplification of cyp6d1, a deltamethrin resistance gene in M. domestica, were determined by high-resolution melting (HRM) analysis and semiquantitative PCR. The accuracy of HRM analysis was validated by sequencing of PCR products. Results The mRNA expression of cyp6d1 in deltamethrin-resistant strain was 50.7-fold that in deltamethrin-sensitive strain, showing significant difference between the two strains (P=0.0002). The PCR products were classified into three genotypes, type A (Tm=85.5℃), type B (Tm=87.3℃), and type C (Tm=88.5℃), according to the mutation analysis of cyp6d1 by HRM. The results from HRM corresponded to the sequences of the three enotypes. As for the genotypes, type A was homozygous 1087G, 1101T, and 1155A (GenBank No. U22367.1), type C was homozygous 1087A, 1101G, and 1155G, and type B was heterozygous for type A and type C. Bioinformatic analysis revealed that this mutation was non-sense mutation. Conclusion The mRNA expression of cyp6d1 is significantly increased in the deltamethrin-resistant strain of M. domestica in Hangzhou, China, which may be one of the mechanisms of deltamethrin resistance increase in M. domestica.

Key words: High-resolution melting, Musca domestica, Deltamethrin resistance, Gene mutation, cyp6d1