中国兽医科学2024，Vol.54Issue(2)：169-173,5.DOI:10.16656/j.issn.1673-4696.2024.0043
鸽圆环病毒TaqMan荧光定量PCR检测方法的建立及初步应用
Establishment and application of PCR assay for detect of pigeon circovirus
郭学波 1赵承锋 2吴瑶 1吴琼 1胡学文1
作者信息
- 1. 汉唐和元(武汉)科技有限公司,湖北武汉 430070
- 2. 安陆市动物疫病预防控制中心,湖北孝感 432600
- 折叠
摘要
Abstract
To establish a method for the rapid and sensitive detection of pigeon circovirus(PiCV),a genome-wide recombinant plasmid of PiCV had been constructed according to a PiCV popular strain genome sequence(GenBank:OL901205.1).Subsequently,multiple sets of primers and probes were designed based on the conserved sequence of PiCV Rep gene.After screening,the optimized primers and probes com-bination which presented the best amplification efficiency was selected.And the TaqMan fluorescent quantitative PCR detection method was established.To test the sensitivity of this method,the synthet-ic standard recombinant plasmid was employed.The results showed that the minimum detection limit of template is 21 copies/μL and the data can keep a good linear relationship over the range 2.1 × 101-2.1 × 108 copies/μL with the linear correlation coefficient R2=0.999 1.This method also show a highly specificity to PiCV which has no signal can be detected from pigeon herpesvirus type 1,pigeon aden-ovirus,Newcastle disease virus and pigeon rotavirus.The repeatability of the method is also excel-lent.The coefficient of variation is less than 2％,and the intra-assay coefficient of variation is less than l％.By using 50 clinical samples,the positive detection rate of this method is 94％while that of PCR is 82％.The coincidence rate of the two methods was 100％.In summary,the TaqMan fluorescent quantitative PCR detection method of PiCV established in this experiment can be used for clinical de-tection of PiCV.关键词
鸽圆环病毒/TaqMan荧光定量PCR/检测/应用Key words
pigeon circovirus/TaqMan fluorescence PCR/detection/application分类
农业科技引用本文复制引用
郭学波,赵承锋,吴瑶,吴琼,胡学文..鸽圆环病毒TaqMan荧光定量PCR检测方法的建立及初步应用[J].中国兽医科学,2024,54(2):169-173,5.
