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建立一种快速、准确鉴别铁皮石斛加工制品的方法。该研究通过对Gen Bank数据库收录的铁皮石斛及其同属近缘物种的ITS序列进行对比分析,根据变异位点设计双阻滞位点特异性鉴别引物,并优化PCR条件,对铁皮石斛及其69种近缘种共362个样品进行扩增和检测。结果显示,在退火温度为60℃的同一PCR反应中,铁皮石斛均扩增出397 bp条带,而其他69种近缘物种均无条带,且该方法灵敏度高,检出限可达0.03 mg·L~(-1)。该文所建立的位点特异性PCR方法可实现铁皮石斛的准确鉴别,具有操作简便、稳定可靠、应用范围广的优点。
To establish a rapid and accurate identification of Dendrobium processed products. In this study, we compared the ITS sequences of Dendrobium officinale and its related species in Gen Bank database, designed double-locus-specific primers and optimized PCR conditions, A total of 362 species of related species were amplified and tested. The results showed that in the same PCR reaction with annealing temperature of 60 ℃, Dendrobium candidum all amplified 397 bp bands, while other 69 species had no bands, and the method was sensitive and the detection limit was up to 0.03 mg · L ~ (-1). The site-specific PCR method established in this paper can achieve the accurate identification of Dendrobium candidum, with the advantages of simple operation, stable and reliable application and wide range of applications.