冬虫夏草实时荧光PCR检测方法的建立

摘要

An efifcient and sensitive method was established to detect Cordyceps sinensis (Berk.) Saccardo using real-time PCR. The internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) in Cordyceps sinensis (Berk.) Saccardo. was used as a target molecular marker and sequence homology analysis was carried out on various Cordyceps species. A set of speciifc primer and probe for real-time PCR was designed which allowed the ampliifcation of 80-bp DNA fragments. The detectable DNA template concentration of the real-time PCR method for Cordyceps sinensis (Berk.) Saccardo was as low as 0.0001 ng/μL.%建立冬虫夏草高效、灵敏的分子检测方法。以冬虫夏草Cordyceps sinensis (Berk.) Saccardo核糖体(rDNA)内转录间隔区(ITS)为分子标记片段，通过对不同虫草同源序列的比对分析，设计对冬虫夏草具有特异性反应的实时荧光PCR引物和探针。该引物检测的目标DNA长度为80bp，对冬虫夏草DNA模版的检测含量可低至0.0001ng/μL。