Loop-mediated isothermal amplification for detection of porcine circovirus type 2
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- 作者：Shun Zhou (1)
Si Han (1) (2)
Jianli Shi (2)
Jiaqiang Wu (2)
Xiaoyuan Yuan (2) (3)
Xiaoyan Cong (2)
Shaojian Xu (2)
Xiaoyan Wu (1) (2)
Jun Li (2) (3)
Jinbao Wang (2) (3) - 关键词：Porcine circovirus type 2 (PCV2) ; Loop ; mediated isothermal amplification (LAMP) ; Rapid detection
- 刊名：Virology Journal
- 出版年：2011
- 出版时间：December 2011
- 年：2011
- 卷：8
- 期：1
- 全文大小：391KB
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Si Han (1) (2)
Jianli Shi (2)
Jiaqiang Wu (2)
Xiaoyuan Yuan (2) (3)
Xiaoyan Cong (2)
Shaojian Xu (2)
Xiaoyan Wu (1) (2)
Jun Li (2) (3)
Jinbao Wang (2) (3)
1. Qingdao Agricultural University, Qingdao, 266109, China
2. Division of Swine Diseases, Shandong Provincial Key Laboratory of Animal Disease Control & Breeding, Institute of Animal Science and Veterinary Medicine Shandong Academy of Agricultural Sciences, Jinan, 250100, China
3. College of Life Sciences, Key Laboratory of Animal Resistance of Shandong Province, Shandong Normal University, Jinan, 250014, China - ISSN：1743-422X
文摘
Background Porcine circovirus type 2 (PCV2) is the primary causative agent of the emerging swine disease known as postweaning multisystemic wasting syndrome (PMWS). Nowadays, polymerase chain reaction (PCR) is still the most widespread technique in pathogen detection. Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method developed in 2000, will possibly replace PCR in the field of detection. To establish a LAMP method for rapid detection of PCV2, two pairs of primers were designed specially from the open reading frame 2 (ORF2) sequences of PCV2. A LAMP method for rapid detection of PCV2 was established. To compare with PCR, sensitivity and specificity of LAMP were evaluated using the optimized reaction system. The LAMP products could be determined by agarose gel electrophoresis or adding SYBR Green I dye. Results The amplification of LAMP could be obtained at 63°C for 60 min. The detection limit was nearly 1 copy of DNA plasmid, more sensitive than PCR. There was no cross-reaction with porcine circovirus type 1 (PCV1), porcine pseudorabies virus (PRV) and porcine parvovirus (PPV) under the same conditions. Conclusions LAMP is an useful rapid detection method with high sensitivity and specificity for PCV2.