摘要
旨在建立川贝母的快速真伪鉴别方法。比较分析川贝母保守性序列及其常见伪品的序列差异,设计了3套LAMP引物,评价了其特异性和灵敏度;利用该技术对市售川贝母样品进行检测,同时用《中华人民共和国药典》的聚合酶链式反应-限制性片段长度多态性法(PCR-RFLP)进行验证。结果表明,三套LAMP引物中,只有加了碱基突变的Set3的引物特异性强;使用Set3引物建立的川贝母LAMP检测方法在川贝母、暗紫贝母和太白贝母样品中显示阳性,在伊贝母、浙贝母、湖北贝母、平贝母等7种常见混淆品中显示阴性,方法特异性良好;LAMP检测的灵敏度在DNA水平上达到32μg/L。采用此LAMP方法进行了30份市售川贝母样品的鉴定,本法实验结果与药典收录的PCR-RFLP法实验结果一致。建立的川贝母LAMP快速鉴别法适用于川贝母及其常见伪品的真伪鉴别。
To developed rapid visual identification method for Fritillaria cirrhosa by loop-mediated isothermal amplification.Methods:According to the DNA sequence of the target,three sets of primers were designed and screened by specificity.Then,specificity and sensitivity of the method was validated.At last,a pair of screened primers were used to identify the commercial Fritillariae cirrhosae samples.And the results were verified by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)method adopted in Chinese Pharmacopoeia(2015 edition).The LAMP detection method established only by using primer set3 showed best performance on amplification specificity.Possitive results could be observed in 3 kinds of Fritillariae,and there was no amplification product in 7 common adulterants.It demonstrated that this LAMP assay was specific.The detection limit of the LAMP method was 32 pg/μL with genomic DNA.Thirty commercial Fritillariae cirrhosae samples,twenty-nine was identified to be positive by this LAMP method,which was consistent with the PCR-RFLP method.The result demonstrated that the specific LAMP assay can be used for rapid authentication of Fritillaria cirrhosa.
作者
徐晓可
邓华明
余至兴
黄逸仪
徐发莲
梁惠倩
黄启红
Xu Xiaoke;Deng Huangming;Yu Zhixing;Huang Yiyi;Xu Falian;Liang Huiqian;Huang Qihong(Lingnan Institute of Technology,Guangzhou 510663,China;Institute of Analysis,Guangdong Academy of Sciences,Guangzhou 510070,China)
出处
《云南化工》
CAS
2022年第12期15-17,共3页
Yunnan Chemical Technology
基金
广东省科技创新战略专项资金项目(大学生科技创新培育)(pdjh2021b1015)。
