安徽农业科学2024，Vol.52Issue(21) ：163-167.DOI:10.3969/j.issn.0517-6611.2024.21.034

微滴式数字PCR技术在菜豆晕疫病菌检测中的应用

Application of Droplet Digital PCR in the Detection of Pseudomonas savastanoi

刘晓宇 钱茱希 郭静 杨静 李彬 吴翠萍 王振华 许剑涛
安徽农业科学2024，Vol.52Issue(21) ：163-167.DOI:10.3969/j.issn.0517-6611.2024.21.034
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  • 万方数据

微滴式数字PCR技术在菜豆晕疫病菌检测中的应用

Application of Droplet Digital PCR in the Detection of Pseudomonas savastanoi

刘晓宇 1钱茱希 1郭静 1杨静 1李彬 1吴翠萍 1王振华 2许剑涛1
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作者信息

  • 1. 南京海关动植物与食品检测中心,江苏 南京 210019
  • 2. 武汉海关技术中心,湖北 武汉 361026
  • 折叠

摘要

为了建立菜豆晕疫病菌(Pseudomonas savastanoi pv.phaseolicola)微滴式数字PCR(ddPCR)检测方法,实现菜豆晕疫病菌的高效率、高精度的检疫鉴定和疫情监控,以菜豆晕疫病菌为研究对象,以位点特异性重组酶作为靶标基因,SSRP_F、SSRP_R、SSRP_P为特异性引物与探针,建立菜豆晕疫病菌ddPCR反应体系,结果表明,菜豆晕疫病菌的ddPCR检测体系的绝对定量检测低限为0.6 copies/μL,ddPCR的检测灵敏度比常规PCR高2个数量级.同时研究了种子携带菜豆晕疫病菌是否具有活性,提高了疫情检出率.

Abstract

In order to realize high efficiency and high precision quarantine identification and epidemic monitoring of Pseudomonas savastanoi pv.phaseolicola(Psp),the detection method of Psp by droplet digital PCR(ddPCR)was established.The ddPCR reaction system was conduc-ted with Psp as research object and site-specific recombinases were used as gene target with SSRP-F/SSRP-R/SSRP-P as primers and probe.The results showed that the detection limit of genomic DNA by ddPCR was 0.6 copies/μL.The detection sensitivity of ddPCR could increase two orders of magnitude than convention PCR.Meanwhile,the activity of Psp carried in peas was studied,which improved the epidemic detec-tion rate.

关键词

微滴式数字PCR技术/菜豆晕疫病菌/快速检测

Key words

Droplet digital PCR/Pseudomonas savastanoi pv.phaseolicola/Rapid detection

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出版年

2024
安徽农业科学
安徽省农业科学院

安徽农业科学

影响因子：0.413
ISSN：0517-6611
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