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猪圆环病毒3型实时荧光定量PCR方法的建立和应用

Development and application of a real-time PCR for the detection of porcine circovirus 3
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摘要 猪圆环病毒3型(PCV3)是近两年报道的一种新病毒,为实现快速检测PCV3的目的,笔者以我国PCV3全基因组为模板设计合成了1对引物和1条TaqMan探针,建立了检测PCV3的实时荧光定量PCR方法。经过标准曲线分析、敏感性、特异性和重复性等一系列试验,表明该方法具有特异性强、敏感性高以及重复性好的特点。试验结果表明,阳性PCV3标准品DNA稀释10-8后(浓度为50 copies/L)仍然可以被检出,不同浓度的组间和组内重复性试验结果显示变异系数均小于3%,最后用36份临床样品进行了应用性评价,结果从中检测出9份阳性样品,表明所建立的方法是一种高效的可用于PCV3的分子诊断技术。 Porcine circovirus 3(PCV3)is one of the novel virus found in pigs since 2016.Diagnosis of PCV3 is difficult with atypical clinical signs.The construct of a real-time PCR was described with a pair of special primers and a Taqman probe targeting a PCV3 genome in the Gen Bank.In order to rapidly detect PCV3,a real-time PCR assay was developed based on the PCV3 genome of the our country isolate,and a pair of primers and a Taqman probe,which were deisgned and synthesized by us.Some tests had proved this assay to be a optimal standard curve linearly with high specifity and sensitivity as well as good reproducibility.It's lowest detectable dilution was 10-8(50 copies/L)in the sensitivity test.For intra-batch and inter-batch reproducibility,the CV values were repeated three times with an average value less than 3%.Additionally,9 positive samples were tested from 36 field samples using this assay.In conclusion,this novel developed real-time PCR assay could be used as an efficient molecular biology tool to facilitate the diagnosis of PCV3.
作者 张志 郝占武 张美晶 吴发兴 刘爽 李晓成 王树双 ZHANG Zhi1, HAO Zhan-wu1,2, ZHANG Mei-jing1, WU Fa-xing1, LIU Shuang1, LI Xiao-cheng1, WANG Shu-shuang1 ( 1. China Animal Health and Epidemiology Center, Qingdao 266032, China; 2. Qingdao Agricultural University, Qingdao 266109, China)
出处 《中国兽医科学》 CSCD 北大核心 2018年第6期686-691,共6页 Veterinary Science in China
基金 科技基础性工作专项(2012FY111000)
关键词 猪圆环病毒3型 实时荧光定量聚合酶链式反应 TAQMAN探针 porcine circovirus 3 (PCV3) real-time PCR TaqMan probe
作者简介 张志(1976-),男,安徽萧县人,副研究员,博士,研究方向为动物病毒学和流行病学,E-mail:zhizh2001@126.com。;通信作者:李晓成(1962-),男,陕西铜川人,研究员,研究方向为动物流行病学,E-mail:lxch20062006@163.com。
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