园艺学报 ›› 2022, Vol. 49 ›› Issue (3): 663-670.doi: 10.16420/j.issn.0513-353x.2020-1027

• 新技术新方法 • 上一篇    下一篇

基于TaqMan MGB探针的山茶叶杯菌实时荧光PCR快速检测方法

吕燕1,2, 段维军1,3,*()   

  1. 1宁波检验检疫科学技术研究院,浙江宁波 315012
    2宁波中盛产品检测有限公司,浙江宁波 315012
    3宁波海关,浙江宁波 315012
  • 收稿日期:2021-06-07 修回日期:2021-11-26 出版日期:2022-03-25 发布日期:2022-03-25
  • 通讯作者: 段维军
  • 基金资助:

TaqMan MGB-based Real-time Fluorescence PCR Method for the Rapid Detection of Ciborinia camelliae

LÜ Yan1,2, DUAN Weijun1,3,*()   

  1. 1Ningbo Academy of Inspection and Quarantine,Ningbo,Zhejiang 315012,China
    2Ningbo Joysun Product Testing Service Company,Ningbo,Zhejiang 315012,China
    3Ningbo Customs District P. R. China,Ningbo,Zhejiang 315012,China
  • Received:2021-06-07 Revised:2021-11-26 Online:2022-03-25 Published:2022-03-25
  • Contact: DUAN Weijun


对山茶叶杯菌(Ciborinia camelliae L. M. Kohn)及其近似种的β-微管蛋白(β-tubulin)基因序列进行了比较分析,设计了1对特异性引物和1条探针,建立了山茶叶杯菌实时荧光PCR检测方法。该方法能特异性检测山茶叶杯菌;体系最佳引物浓度、探针浓度分别为0.8和0.7 μmol · L-1;在10 μL反应体系中,最低检测限可达1.0 pg;该方法可用于疑似携带山茶叶杯菌样品的检测与初筛。

关键词: 山茶, 花腐病, 山茶叶杯菌, TaqMan MGB探针, 实时荧光PCR, 快速检测


Based on β-tubulin gene of Ciborinia camelliae L. M. Kohn and related isolates,a pair of specific primers and a TaqMan MGB probe were designed and synthesized. A novel real-time fluorescent PCR was established to detect C. camelliae. The optimized primer and probe concentrations were 0.8 and 0.7 μmol · L-1,respectively. The minimum detection limit of this method is up to 1.0 pg for 10 μL reaction system. The method was specific,sensitive,easily to operate and fast,which provide a valuable tool for early rapid detection and identification of C. camelliae.

Key words: Camellia, camellia flower blight, Ciborinia camelliae, TaqMan MGB probe, real-time fluorescent PCR, rapid detection