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
蔡彦秋 1查杰 1沐阳 1陈辉1
作者信息
- 1. 泰州市疾病预防控制中心,江苏泰州 225300
- 折叠
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Oligo 7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 5.8×102 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.
Abstract
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 5.8×102 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Key words
Burkholderia gladioli/real-time fluorescent PCR/specificity/sensitivity引用本文复制引用
出版年
2024
NETL
NSTL
万方数据