铁皮石斛PCR鉴定引物的比较和筛选研究

Comparative and Screening Study on PCR Identification Primers for Dendrobium officinale

目的:比较并筛选基于PCR技术的铁皮石斛成分鉴定的高特异性和高灵敏度引物。方法:选取1组专利报道和2组文献报道的铁皮石斛特异性PCR鉴定引物,优化其退火温度,比较其特异性和灵敏度,筛选出高特异性和高灵敏度的引物。结果:3组引物的扩增能力、特异性和灵敏度均存在差异,其中1组以铁皮石斛ITS2区为靶标、引入错配碱基的双阻滞铁皮石斛成分鉴定引物,具有最佳的特异性和灵敏度。结论:通过比较3组铁皮石斛特异性PCR引物,优选出1组铁皮石斛成分鉴定引物,为铁皮石斛成分及其产制品的鉴定提供了参考依据。

Objective To comparie and screen high specificity and sensitivity primers in Dendrobium officinale ingredient identification methods based on PCR.Method PCR identification primers for Dendrobium officinale were selected from one group of patent reports and two groups of literature reports,and their annealing temperatures were optimized.The amplification specificity and sensitivity of the primers were compared,and high specificity and sensitivity primers were screened out.Result There were differences in amplification capacity,specificity and sensitivity respectively among these 3 groups of primers.One group targeting the ITS2 region of Dendrobium officinale and introducing a pair of double-blocked specific primers for Dendrobium officinale identification had the best specificity and sensitivity.Conclusion By comparing 3 groups of Dendrobium officinale-specific PCR primers,one group of primers was selected as a preferred one,which provided a reference for the identification of Dendrobium officinale and its products.