利用实时荧光定量PCR方法快速区分田鼠型和非田鼠型鼠疫耶尔森菌
Using real-time fluorescence quantitative PCR method to rapidly differentiate Microtus and non-Microtus Yersinia pestis
耿朝阳 1王宇萌 2贺金荣 2李伟 2王占黎 3郑霄2
作者信息
- 1. 内蒙古科技大学包头医学院公共卫生学院,内蒙古包头 014000;中国疾病预防控制中心传染病预防控制所
- 2. 中国疾病预防控制中心传染病预防控制所
- 3. 内蒙古科技大学包头医学院公共卫生学院,内蒙古包头 014000
- 折叠
摘要
目的 建立一种快速区分田鼠型和非田鼠型鼠疫耶尔森菌的实时荧光定量PCR方法.方法 鼠疫耶尔森菌天冬氨酸酶基因(aspA)与致病力相关,鼠疫菌aspA基因1087碱基(G/T)等位基因突变能有效区分我国田鼠型与非田鼠型鼠疫菌.本研究基于该等位基因突变位点设计特异性差异探针和引物,发展差异探针荧光定量PCR方法检测田鼠型与非田鼠型鼠疫菌,对内蒙古、四川田鼠型和非田鼠型鼠疫疫源地以及我国其他鼠疫疫源地代表菌株应用评价.结果 建立的特异性差异探针荧光定量PCR方法能准确区分田鼠型鼠疫菌和非田鼠型鼠疫菌,在其他非鼠疫菌上没有特异扩增.结论 此方法可以对田鼠型和非田鼠型鼠疫耶尔森菌进行准确区分,适用于我国鼠疫疫源地分离菌株的快速鉴定.
Abstract
Objective To establish a real-time fluorescence quantitative PCR method for rapid differentiating the microtus and non-microtus Yersinia pestis.Methods The aspartase gene(aspA)of Yersinia pestis is pathogen-related gene,and the allelic gene mutation(G/T)at 1087 base pair site of the asp A gene can effectively distinguish the microtus and non-microtus Yersinia pestis.Based on the allelic gene mutation site,specific differentiating probes and primers were designed to establish a fluorescence quantitative PCR method to identigy microtus and non-microtus Yersinia pestis.The discriminatory effectiveness of this method was evaluated by testing representative strains from plague foci in China.Results The established specific differential probe fluorescence quantitative PCR method can accurately distinguish the microtus and non-microtus Yersinia pestis,with no amplification observed in non-Yersinia pestis strains.Conclusion This method can be used to differentiate the microtus and non-microtus Yersinia pestis strains,making it suitable to identify surveillance strains quickly from plague foci in China.
关键词
鼠疫耶尔森菌/实时荧光定量PCR/aspA基因Key words
Yersinia pestis/Real-time fluorescence quantitative PCR/aspA gene引用本文复制引用
基金项目
"生物安全关键技术研发"重点专项(2021YFC1200200)
内蒙古自治区科技重大专项(2021ZD0006)
出版年
2024
NETL
NSTL
万方数据