冬虫夏草实时荧光PCR检测方法的建立

A Real-Time PCR Method for Detection of Cordyceps sinensis(Berk.) Sacc.

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。

An efficient 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 specific primer and probe for real-time PCR was designed which allowed the amplification 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.