食品中唐菖蒲伯克霍尔德氏菌实时荧光PCR检测的研究  

Research of Burkholderia gladioli in Food by Real-Time Fluorescent PCR Method

作　　者：蔡彦秋[1] 查杰[1] 沐阳 陈辉 CAI Yanqiu;ZHA Jie;MU Yang;CHEN Hui(Taizhou Municipal Center for Disease Control and Prevention,Taizhou 225300,China)

机构地区：[1]泰州市疾病预防控制中心,江苏泰州225300

出　　处：《食品安全导刊》2024年第20期44-47,共4页China Food Safety Magazine

基　　金：泰州市科技计划项目(TS201911);江苏省卫生计生委科研项目(Y2018065)。

摘　　要：目的:建立唐菖蒲伯克霍尔德氏菌的核酸检测方法。方法:根据16S~23S rRNA基因片段序列,利用Oligo7设计TaqMan探针及引物,并验证实时荧光PCR检测方法的特异性和灵敏性。结果:用实时荧光PCR检测方法检测15株标准菌株,只有唐菖蒲伯克霍尔德氏菌呈阳性,验证了此方法具有良好的特异性。灵敏度实验中得出菌液灵敏度为5.8×10~2 CFU·mL^(-1),DNA灵敏度为7.2×10^(-4) ng·μL^(-1)。结论:建立的实时荧光PCR检测方法有良好的特异性和灵敏度,该方法可用于唐菖蒲伯克霍尔德氏菌的快速检测,具有较好的应用价值。Objective:To establish a method for Burkholderia gladioli DNA detection.Method:Based on 16S~23S rRNA gene fragment sequence,TaqMan probes and primers were designed by Oligo7.The method of real-time fluorescent PCR detection was further verified for its specificity and sensitivity.Result:Detecting 15 standard strains by real-time fluorescent PCR detection method,only Burkholderia gladioli was positive.So it’s confirmed that this method has good specificity.Through sensitivity experiments of real-time fluorescent PCR method,it was detected that the sensitivity of the bacterial solution was 5.8×10~2 CFU·mL~(-1) and that of DNA was 7.2×10~(-4) ng·μL~(-1).Conclusion:The established real-time fluorescent PCR method has strong specificity and sensitivity so that it can be applied to rapid detection of Burkholderia gladioli,demonstrating good application value.

关 键 词：唐菖蒲伯克霍尔德氏菌 实时荧光PCR 特异性 灵敏性 

分 类 号：TS207.4[轻工技术与工程—食品科学] O657.3[轻工技术与工程—食品科学与工程]