兰花细菌性褐腐病菌实时荧光PCR快速检测方法的建立  

Establishment of real-time fluorescent PCR method for detection of Erwinia cypripedii

作　　者：季蕾 邵秀玲 杨斌 孙铮 齐振华 厉艳 Ji Lei;Shao Xiuling;Yang Bin;Sun Zheng;Qi Zhenhua;Li Yan(Inspection and Quarantine Technology Center of Qingdao Customs District,,Qingdao 266002,China)

机构地区：[1]青岛海关检验检疫技术中心,山东青岛266002

出　　处：《植物检疫》2023年第6期31-36,共6页Plant Quarantine

基　　金：国家重点研发计划(2021YFD1400100,2021YFD1400103)。

摘　　要：本研究以兰花细菌性褐腐病菌的16S rDNA基因片段为靶标,建立了一套兰花细菌性褐腐病菌的实时荧光PCR快速检测体系。通过对反应条件的优化,确定最佳反应体系的引物终浓度和探针终浓度分别为0.8μmol/L和0.9μmol/L;在10μL反应体系中,最低检出限可达8.61 pg,且具有较好的特异性和重复性。因此,该体系的建立为该病菌的检测、鉴定提供了技术支持。The study has established a real-time fluorescent PCR rapid detection system for E.cypripedi by targeting the 16S rDNA sequence of Erwinia cypripedi.After the optimizing of reaction conditions,the opti-mized primer and probe final concentrations of the method were 0.8μmol/L and 0.9μmol/L,rerspectively.The minimum detection limit of this method is up to 8.61 pg for 10μL reacation system with good specificity and repeatability.Therefore,the establishment of this system provides technical support for the detection and identification of E.cypripedi.

关 键 词：兰花细菌性褐腐病菌 实时荧光PCR 快速检测 

分 类 号：S41[农业科学—植物保护]