应用Proofman-LMTIA技术双重检测当归和东当归

Dual Detection of Angelica sinensis(Oliv.)Diels and Angelica acutiloba(Sieb.et Zucc.)Kitagawa by Proofman-LMTIA Technique

通过梯型熔解温度核酸等温扩增技术(Ladder-shape melting temperature nucleic acid isothermal amplification,LMTIA)与校对酶介导探针(Proofreading enzyme-mediated probe cleavage,Proofman)联合的方法对当归和东当归进行双重检测。以ddH2O为阴性模板,当归和东当归基因组DNA为阳性模板,测定当归和东当归引物的最适温度、灵敏度及稳定性,并进行了真实样品检测。结果表明,应用Proofman-LMTIA双重检测当归和东当归,引物测定的最适温度为62℃,当归引物灵敏度低至10 pg/μL,东当归引物灵敏度低至1 pg/μL,具有良好的稳定性。7份市售当归样品经检测均为当归。该研究可为当归和东当归的鉴别提供一种新的检测方法。

Dual detection of Angelica sinensis and Angelica acutiloba was performed by a combined method of ladder-shape melting temperature nucleic acid isothermal amplification technique(LMTIA)and proofreading enzyme-mediated probe(Proofman).The optimum temperature,sensitivity,and stability of the primers for Angelica sinensis and Angelica acutiloba were determined using distilled water as the negative template and Angelica sinensis and Angelica acutiloba genomic DNA as the positive template,and real samples were tested.It was proved that the Proofman-LMTIA dual detection of Angelica sinensis and Angelica acutiloba had an optimal temperature of 62℃,and the sensitivity of the primers was as low as 10 pg/μL for Angelica sinensis and 1 pg/μL for Angelica acutiloba,with good stability.Seven commercial Angelica sinensis samples were tested and all were identified as Angelica sinensis.This study provides a new detection method for the identification of Angelica sinensis and Angelica acutiloba.