Loop-mediated isothermal amplification for detection of porcine circovirus type 2

文献类型： 外文期刊

第一作者： Han, Si

作者： Han, Si;Shi, Jianli;Wu, Jiaqiang;Yuan, Xiaoyuan;Cong, Xiaoyan;Xu, Shaojian;Wu, Xiaoyan;Li, Jun;Wang, Jinbao;Zhou, Shun;Han, Si;Wu, Xiaoyan;Yuan, Xiaoyuan;Li, Jun;Wang, Jinbao

作者机构：

关键词： Porcine circovirus type 2 (PCV2);Loop-mediated isothermal amplification (LAMP);Rapid detection

期刊名称：VIROLOGY JOURNAL （ 影响因子：4.099； 五年影响因子：3.719 ）

ISSN： 1743-422X

年卷期： 2011 年 8 卷

页码：

收录情况： SCI

摘要： 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 degrees 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.

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