[1]李晓菲,陈 婷,孙爱荣,等.猪圆环病毒3型TaqMan-MGB荧光定量PCR方法的建立及应用[J].中国预防兽医学报,2019,(02):151-155.[doi:0.3969/j.issn.1008-0589.201805012]
 LI Xiao-fei,CHEN Ting,SUN Ai-rong,et al.Establishment and application of TaqMan-MGB real-time PCR assay for detection of porcine circovirus 3[J].Chinese journal of preventive veterinary medicine,2019,(02):151-155.[doi:0.3969/j.issn.1008-0589.201805012]
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猪圆环病毒3型TaqMan-MGB荧光定量PCR方法的建立及应用()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年02
页码:
151-155
栏目:
诊断和检测技术
出版日期:
2019-02-15

文章信息/Info

Title:
Establishment and application of TaqMan-MGB real-time PCR assay for detection of porcine circovirus 3
文章编号:
1008-0589(2019)02-0151-05
作者:
 

李晓菲陈 婷孙爱荣葛晓杰黄 艳徐 婷王彩宏魏笑笑秦立廷*

 (山东新希望六和集团有限公司,山东 青岛 266000)
Author(s):
 

LI Xiao-fei CHEN Ting SUN Ai-rong GE Xiao-jie HUANG Yan XU Ting WANG Cai-hongWEI Xiao-xiao QIN Li-ting*

 (Shandong New Hope Liuhe Group Co., Ltd, Qingdao 266000, China)
关键词:
猪圆环病毒3型TaqMan-MGB荧光定量方法临床样品
Keywords:
 porcine circovirus 3  TaqMan-MGB real-time PCR assay  clinical samples
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201805012
文献标志码:
A
摘要:
为建立猪圆环病毒3型(PCV3)荧光定量PCR检测方法,本研究根据GenBank中PCV3基因序列设计特异性引物和探针,经过反应体系和条件优化,建立了特异性检测PCV3的TaqMan-MGB荧光定量PCR方法。该检测方法在4.78×101拷贝/μL~4.78×109拷贝/μL质粒标准品范围内均有良好的线性关系;该方法特异性试验结果显示,其与多种常见猪病病毒均无交叉反应,特异性良好;本研究建立的方法敏感性是常规PCR方法的100倍,敏感性较高;批内批间重复性试验变异系数均小于2.3 %,重复性良好。对临床样品的检测结果显示,该方法对PCV3的检出率高于常规PCR方法,并且PCV3阳性样品多存在混合感染情况。该方法的建立为PCV3的实验室诊断及流行病学调查提供了快速、准确的检测手段。
 
Abstract:
To established a sensitive detection method of porcine circovirus 3 (PCV3), a TaqMan-MGB real-time PCR assay was established using specific primers and probe designed according to the genome sequence of PCV3. With the optimized reactions conditions, the established method was specific for PCV3 detection without cross-reaction with other common porcine viruses. Results showed that the method had linearity within the template ranges from 4.78×101 to 4.78×109 copies/μL and was 100 times more sensitive than that of routine PCR assay. The coefficient of variation in the reproducible assays was less than 2.3%. The positive detection rate of PCV3 in clinical samples using the developed method was higher than that using the conventional PCR method and the PCV3 positive samples were mostly mixed infection. The establishment of the real-time PCR method provides a rapid and accurate detection means for the laboratory diagnosis and epidemiological investigation of PCV3.

参考文献/References:

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                               (本文编辑:彭永刚)

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更新日期/Last Update: 2019-03-06