Development of a Real-time PCR Method for Detection of Type 2 Porcine Circovirus

FENG Zhi-xin; JIANG Ping*; WANG Xian-wei; LI Yu-feng; XU Jia-rong

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August 2006

FENG Zhi-xin, JIANG Ping*, WANG Xian-wei, LI Yu-feng and XU Jia-rong. Development of a Real-time PCR Method for Detection of Type 2 Porcine Circovirus[J]. Virologica Sinica, 2006, 21(4): 371-374.

Citation: FENG Zhi-xin, JIANG Ping*, WANG Xian-wei, LI Yu-feng, XU Jia-rong. Development of a Real-time PCR Method for Detection of Type 2 Porcine Circovirus .VIROLOGICA SINICA, 2006, 21(4) : 371-374.

猪圆环病毒2型TaqMan实时PCR检测方法的建立

1.
南京农业大学农业部动物疫病诊断与免疫重点开放实验室，江苏南京，210095

摘要

设计合成了一套引物和TaqMan探针，特异性扩增猪圆环病毒2型（PCV2）ORF2基因，在国内首次建立了快速定量检测PCV2的实时PCR方法，且该方法具有较好的特异性和重复性，对PCV2DNA检测下限为1copy/μL ，敏感性比常规PCR高106倍；分别用该法和普通PCR方法对PMWS人工发病猪的10份组织及30份血清样品检测，结果表明该方法具有更快速、灵敏、准确、低污染等优点，并可以对PMWS的早期检测、预防起到指示作用。
- 猪圆环病毒2型（PCV2）
- , TaqMan
- , 实时PCR

Development of a Real-time PCR Method for Detection of Type 2 Porcine Circovirus

1.
Key Laboratory of Animal Disease Diagnostic and Immunology, Ministry of Agricultur, Nanjing Agricultural University，Nanjing 210095，China

Abstract

One set of primers and a TaqMan probe were designed based on the nucleotide sequence within ORF2 of Porcine circovirus type 2 (PCV2). A quantitative TaqMan real-time PCR was developed for detecting PCV2 with highly specificity, reproducibility and more sensitivity than conventional PCR. Standard dilutions allowed absolute quantification of the amount of viral DNA to 1copy/μL, which is 106 times more sensitive than conventional PCR. The result of detecting PCV2 from tissues and sera of pigs with PMWS also confirms the high sensitivity and specificity of this method. This method is probably a powerful enough tool for early diagnosis of PMWS.
- PCV2
- , TaqMan
- , Real-time PCR.

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Development of a Real-time PCR Method for Detection of Type 2 Porcine Circovirus

1. Key Laboratory of Animal Disease Diagnostic and Immunology, Ministry of Agricultur, Nanjing Agricultural University，Nanjing 210095，China

Abstract: One set of primers and a TaqMan probe were designed based on the nucleotide sequence within ORF2 of Porcine circovirus type 2 (PCV2). A quantitative TaqMan real-time PCR was developed for detecting PCV2 with highly specificity, reproducibility and more sensitivity than conventional PCR. Standard dilutions allowed absolute quantification of the amount of viral DNA to 1copy/μL, which is 106 times more sensitive than conventional PCR. The result of detecting PCV2 from tissues and sera of pigs with PMWS also confirms the high sensitivity and specificity of this method. This method is probably a powerful enough tool for early diagnosis of PMWS.