中国计量大学学报2024，Vol.35Issue(2)：357-362,6.DOI:10.3969/j.issn.2096-2835.2024.02.020
铁皮石斛实时荧光RPA快速鉴别方法的建立
Establishment of a real-time fluorescent recombinase polymerase amplification(RPA)for the Dendrobium officinale
摘要
Abstract
Aims:This paper aims to establish a rapid identification method for Dendrobium officinale based on recombinase polymerase amplification(RPA).Methods:Assemblies of primers and probes targeting the rDNA internal transcribed spacer(ITS)region of D.officinale were designed and screened.The real-time RPA reaction condition was optimized,while the specificity,sensitivity and stability of the test were determined.Results:D.officinale could be specifically detected with 15 min at 39 ℃ using the established real-time fluorescent RPA method.No cross reactions were recorded with the DNA of D.monili forme,D.loddigesii,D.hercoglossum,D.devonianum and D.aduncum.The detection limit for the target DNA was 1.60×10-3 ng/μL with a high stability.Conclusions:The established real-time fluorescence RPA method is simple and rapid,and has high specificity,sensitivity and stability,which is applicable for rapid and accurate identification of D.officinale.关键词
铁皮石斛/重组酶聚合酶扩增/荧光/快速鉴别Key words
Dendrobium officinale/recombinase polymerase amplification/fluorescence/rapid identification分类
生物科学引用本文复制引用
赵立佳,邵珠龙,王正亮,俞晓平..铁皮石斛实时荧光RPA快速鉴别方法的建立[J].中国计量大学学报,2024,35(2):357-362,6.基金项目
浙江省属高校基本科研业务费专项项目(No.2022YW82) （No.2022YW82）
