猪流行性腹泻病毒RT-LAMP检测方法的建立与应用

发布时间：2018-03-24 01:24

本文选题：猪流行性腹泻病毒　切入点：RT-LAMP　出处：《中国兽医学报》2017年07期

【摘要】：针对猪流行性腹泻病毒(PEDV)的M基因设计LAMP引物,对反应体系的条件优化并进行特异性及敏感性等试验。结果显示:该方法在60℃下恒温扩增60min,使PEDV的M基因得到了高效率特异性扩增。本试验方法特异性好,与猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRSSV)、猪圆环病毒2型(PCV-2)、猪细小病毒(PPV)及猪伪狂犬病病毒(PRV)等无交叉反应,同时也具有较好的灵敏性,最低检测分子拷贝数为1.0×102 copies/μL。反应结束后加入SYBR GreenⅠ在紫外灯下观察颜色变化,结果显示阳性扩增产物呈现绿色荧光。结果表明:该LAMP检测方法特异性强,灵敏度高,操作便捷,适于临床检测PEDV。
[Abstract]:The porcine epidemic diarrhea virus (PEDV) M gene LAMP primers were designed on the reaction system and conditions were optimized specificity and sensitivity tests. The results show that the method of constant temperature at 60 DEG C of 60min amplification, M gene PEDV was amplified. The efficiency test method with good specificity. Classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRSSV), porcine circovirus type 2 (PCV-2), porcine parvovirus (PPV) and pseudorabies virus (PRV) and no cross reaction, but also has good sensitivity, lowest molecular detection of copy number after joining SYBR Green I observed the color change in the UV lamp is 1 x 102 copies/ L. reaction, the results showed that the positive PCR products showed green fluorescence. The results show that the LAMP method has strong specificity, high sensitivity, convenient operation, suitable for clinical detection of PEDV.

【作者单位】：贵州大学动物科学学院;贵州省动物疫病预防控制中心;贵州省贵阳市农业委员会;贵州省生猪质量安全工程技术研究中心;贵州省畜禽遗传资源管理站;
【基金】：贵州省科学技术厅农业攻关资助项目(黔科合NY[2015]3009-1号) 贵州省生猪质量安全工程技术研究中心建设资助项目(黔科合农C字[2011]4022号) 贵州省动物疫病防控与兽医公共卫生保障科技创新人才团队资助项目(黔科合人才团队[2015]4016号) 贵州省农业委员会科技计划资助项目(2015-01号)
【分类号】：S852.651

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