药用昆虫神农洁蜣螂炮制品DNA条形码序列的测定与分析
DNA barcoding analysis of processed medicinal insect Catharsius molossus
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摘要: 当药用昆虫被加工炮制成碎块或粉末状态时,其昆虫类别和真伪的鉴别就显得十分困难.未经准确鉴定的药用昆虫会影响临床用药安全.为寻找准确、灵敏且快速的检测方法,采用DNA条形码技术, 分别以药用神农洁蜣螂 (Catharsius molossus) 炮制品药材的完整和破碎个体为研究材料, 对其mtCOI基因进行提取、扩增及测序分析, 探讨DNA条形码技术对炮制后药用蜣螂鉴定的可行性.通过对不同DNA提取方法的比较, 成功从炮制后干燥药材虫体内提取出总DNA并扩增出mtCOI基因片段.测序分析结果表明mtCOI基因序列能够区分神农洁蜣螂和其形态近似种.作为药用神农洁蜣螂DNA条形码标准片段的mtCOI基因序列已投递至GenBank.研究还表明, 碎块状药用蜣螂药材中混杂有一定比例的其他昆虫.该结果有助于建立适合加工炮制后药用蜣螂的DNA条形码分子鉴定标准, 并为保证临床用药安全提供支持.Abstract: The identification of medicinal insects is a complex task, especially when they are processed into pieces or powders. This difficulty has potential to create severe complications. For example, inaccurate identification can affect the safety of clinical application for corresponding medicinal insects. A quick and accurate method to identify these kinds of organisms is needed. Here, we amplified and sequenced the mtCOI gene in processed product of Catharsius molossus, including intact individuals and broken individuals, to test the feasibility of DNA barcoding this kind of sample. After comparing results of different DNA extraction methods, we finally succeed in amplifying and sequencing the barcoding segments in these samples. Our method’s barcoding sequences could clearly distinguish between C. molossus and its allied species. The data also indicated that a degree of interfusion of other insects was present in the broken medicinal C. molossus product. These results are quite promising to the establishment of performance criteria for future identification of medicinal C.molossus that will help ensure the safety application of these medicinal insects.