金钱松实时荧光PCR鉴定方法的建立

A real-time PCR for identification of Pseudolarix amabilis

本研究根据金钱松核糖体基因的内转录间隔区(ITS)设计特异性引物与荧光探针,建立了金钱松的实时荧光PCR检测方法。利用该方法检测4个不同来源的金钱松样本,均能够得到典型的扩增曲线,与53个松柏科其他树种如日本五针松、罗汉松、湿地松样本无交叉反应,表明了该方法的特异性。灵敏度可达1.6×10^-5ng/μL。该方法可快速、灵敏、特异地鉴定金钱松树种,适用于口岸进出口苗木的鉴定。

A real-time PCR method for identification of Pseudolarix amabilis was developed with specific primers and probe designed from the internal transcribed spacer of Pseudolarix amabilis.Specificity study showed that typical amplification curves could be obtained for 4 Pseudolarix amabilis samples,but not for 53 other samples such as Pinus parviflora,Podocarpus macrophyllus,Pinus elliottii.Sensitivity of this method was 1.6×10^-5 ng/μL.The established real-time PCR technique in this study enables rapid,sensitive and specific identification of Pseudolarix amabilis,which is suitable for the application in import and export of seedlings surveillance.