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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。
LAMP primers were designed for the M gene of porcine epidemic diarrhea virus (PEDV), and the conditions of the reaction system were optimized and tested for specificity and sensitivity. The results showed that this method was carried out by constant temperature amplification at 60 ℃ for 60min, which made M gene of PEDV highly efficient and specific amplification. The test method has good specificity and is highly specific to CSFV, PRRSV, PCV-2, PPV and PRV PRV) and so no cross-reaction, but also has good sensitivity, the minimum copy number of detection molecules 1.0 × 102 copies / μL. After the reaction, SYBR Green I was added to observe the color change under UV light. The results showed that the positive amplification product showed green fluorescence. The results showed that the LAMP detection method is specific, sensitive and easy to operate. It is suitable for clinical detection of PEDV.