期刊论文

Establishment and application of Real-Time Fluorescence Quantitative PCR Assay for Chicken Escherichia Coli

中文

湖南畜牧兽医

1006-4907

2024

1

31-35

10.3969/j.issn.1006-4907.2024.01.011

:湖南现代农业产业技术体系项目

本校为其他机构

为建立一种灵敏、准确且快速检测禽源大肠杆菌的方法,根据禽源大肠杆菌uidA基因的一节特异性保守序列设计引物,建立用于禽源大肠杆菌的荧光定量PCR(qPCR)检验方法,并对其各方面进行评价.试验数据表明所建立的qPCR方法的Ct值与标准品在4.69 × 102～4.69×109 copies/μL范围内存在优良的线性关系,线性相关系数为R2=0.993;该方法标准曲线方程为y=-3.2786x+39.032,熔解曲线表现为单峰,不存在非特异性扩增.对重组质粒标准品的最低检测浓度为4.69 × 102 copies/μL,是普通PCR方法的1000倍.该方法检测临床样本阳性率为68.3％(41/60),普通PCR阳性检出率为23.3％(14/60),阳性检出率高出45％.此次研究建...

Aimto establish a sensitive,precise and rapid Real-time fluorescence quantitative PCR means for the detection of chicken escherichia coli,we designed primers based on a specific conserved sequence of uidA gene,which is chicken escherichia coli's specific gene.The uidA gene of Avian Pathogenic Escherichia Coli was constructed on 19T vector.We utilized the recombinant plasmid as a standard positive template to set up means for the detection of chicken escherichia coli and evaluated the specificity,Sensitiveness and repeatability of the method.The...