【目的】 DNA条形码技术已经在多个类群中得到了广泛应用，但对数量巨大的鳞翅目昆虫而言，仍然缺失大量数据，尤其是形态鉴定较为困难的小蛾类和很多中型蛾类，尚无法构建较为完善的DNA条形码系统。本研究旨在为鳞翅目害虫DNA条形码系统的构建和完善提供数据来源及支撑，验证COⅠ基因作为DNA条形码通用基因的准确性，探讨28S rDNA的D2基因片段作为DNA条形码辅助基因的可行性，并检验目前BOLD系统的鉴定成功率。【方法】 对采集自北京白羊沟风景区的小蛾类和中型蛾类490头标本进行形态鉴定和DNA测序，分别基于COⅠ及28S D2基因计算种内种间遗传距离，并构建了NJ系统发育树。【结果】 BOLD系统的鉴定成功率为65%，对小蛾类和夜蛾类鉴定成功率较低。基于COⅠ基因的NJ树鉴定成功率为94.4%，基于28S D2基因的NJ树鉴定成功率为89.4%。【结论】 结合种内与种间遗传距离结果，COⅠ基因适合作为鳞翅目蛾类DNA条形码通用基因，28S D2基因较为保守，不适合作为DNA条形码的辅助基因。

[Objectives]  DNA barcoding has been widely applied to many insect taxa but not a large number of the Lepidoptera, especially smaller and medium-sized moths, which are difficult to identify morphologically. It is not yet possible to build a complete DNA barcoding system. This study aims to provide data sources and support for the construction and improvement of DNA barcoding systems for lepidopteran pests, verify the accuracy of the COⅠgene as a DNA barcoding gene, explore the feasibility of using the 28S D2 gene as a DNA barcoding assistant gene, and test the success rate of BOLD system identification. [Methods]  Morphological identification and DNA sequencing of 490 small and medium-sized moths collected in the Baiyanggou Scenic Area in Beijing were carried out. Intraspecific and interspecific distances were then calculated based on differences in the COⅠand 28S D2 genes and NJ phylogenetic trees were constructed. [Results]  The BOLD system had an average success rate of 65% but the success rate for smaller moths and noctuids was low. NJ trees based on the COⅠand the 28S D2 genes successfully identified 94.4% and 89.4% of specimens, respectively. [Conclusion]  The COⅠgene is suitable for general DNA barcoding of moths whereas the 28S D2 gene is more conservative and therefore not suitable as an assistant gene for DNA barcoding.