动物医学进展2025,Vol.46Issue(1) :68-73.

鸽圆环病毒荧光定量PCR检测方法的建立及应用

Application and Establishment of a Real-time PCR Detection Method for Pigeon Circovirus

刘存 刘砚涵 祝夕超 李法凯 张栋 邵新宇 田野 孙圣福 陈静
动物医学进展2025,Vol.46Issue(1) :68-73.

鸽圆环病毒荧光定量PCR检测方法的建立及应用

Application and Establishment of a Real-time PCR Detection Method for Pigeon Circovirus

刘存 1刘砚涵 1祝夕超 2李法凯 1张栋 1邵新宇 1田野 3孙圣福 1陈静1
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作者信息

  • 1. 山东省动物疫病预防与控制中心(山东省人畜共患病流调监测中心),山东济南 250100
  • 2. 莱州市程郭畜牧兽医站,山东烟台 261437
  • 3. 山东省农业科学院家禽研究所,山东济南 250023
  • 折叠

摘要

为了建立鸽圆环病毒(PiCV)的快速检测方法,依据PiCV Rep基因的保守序列区域设计了荧光定量PCR的引物和探针,并建立了快速检测PiCV的荧光定量PCR方法.结果显示,构建的PiCV荧光定量PCR方法在1.44×10,1~1.44×107/μL拷贝标准品间具有良好的线性关系,线性相关系数达0.986 6;该方法具有良好的敏感性,其检测病毒系列含量下限为14.4拷贝/μL;该方法特异性良好,与鸽新城疫病毒、鸽疱疹病毒、鸽腺病毒等病毒及鸽组织DNA不存在交叉反应;该方法重复性较好,批内重复试验变异系数均介于0.454%~0.473%,批间重复试验变异系数均介于0.367%~0.869%.与传统普通PCR方法相比,所建立的PiCV荧光定量PCR方法敏感性更高,临床样品PiCV检出率更高,两者检测符合率为93.75%.结果表明,所建立的PiCV荧光定量PCR方法为临床检测鸽圆环病毒提供了一种快速、灵敏的检测方法,为PiCV的流行病学调查和主动监测提供了技术支撑.

Abstract

To develop a rapid diagnostic method for pigeon circovirus(PiCV),a real-time PCR assay method for PiCV was established,using the primers and probe designed on the conserved sequence of Rep gene.The standard curve of the real-time PCR assay had good linear relationships in the range of 1.44X 101-1.44×107 copies/μL of template with the linear correlation coefficients to 0.986 6.The assay had a sensi-tivity of 1.44×10 copies/μL which showed that it had a good sensitivity.There was no cross-reaction with pigeon Newcastle disease virus,pigeon herpesvirus,pigeon adenovirus,pigeon salmonellosis and pigeon tis-sue DNA.This method has good repeatability,with coefficients of valuation ranging from 0.454%to 0.473%for intrabatch repeated experiments and 0.367%to 0.869%for interbatch repeated experiments.Compared with traditional PCR assay,the real-time PCR assay established in this study had better sensitiv-ity and slightly higher detection rate in clinical samples,with a detection coincidence rate 93.75%.In con-clusion,the real-time PCR assay performed as a diagnosis tool for PiCV detection,which provides a techni-cal support for the epidemiological investigation of PiCV.

关键词

鸽圆环病毒/荧光定量PCR/Rep基因/检测

Key words

Pigeon circovirus/Real-time PCR/Rep gene/Detection method

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出版年

2025
动物医学进展
西北农林科技大学

动物医学进展

北大核心
影响因子:0.857
ISSN:1007-5038
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