猪繁殖与呼吸综合征病毒RT-LAMP快速检测方法的建立
发布时间：2018-08-01 12:38
【摘要】：为建立一种快速、灵敏、简便的猪繁殖与呼吸综合征病毒(PRRSV)早期检测方法,本研究利用逆转录环介导恒温扩增技术(RT-LAMP),针对PRRSV的ORF5基因片段设计了4条引物,利用Bst DNA聚合酶在65℃恒温条件下进行逆转录扩增,通过1.0%琼脂糖凝胶电泳和加入SYBR GreenⅠ染料肉眼判断结果,建立了PRRSV RT-LAMP检测方法。结果表明,该检测方法具有良好的特异性,与其他常见病毒如猪瘟病毒、猪圆环病毒2型、猪细小病毒、猪伪狂犬病病毒等无交叉反应,较普通RT-PCR灵敏性高100倍。采用RT-LAMP和RT-PCR分别对10份临床样本同时进行检测,符合率为100%。因此,本研究建立的RT-LAMP是一种可适用于临床PRRSV检测的快速、简单、灵敏、特异的检测方法。
[Abstract]:In order to establish a rapid, sensitive and simple method for early detection of porcine reproductive and respiratory syndrome virus (PRRSV), four primers were designed for ORF5 gene fragment of PRRSV by reverse transcription-loop mediated isothermal amplification (RT-LAMP). The Bst DNA polymerase was amplified by reverse transcriptase at 65 鈩，
本文编号：2157608
[Abstract]:In order to establish a rapid, sensitive and simple method for early detection of porcine reproductive and respiratory syndrome virus (PRRSV), four primers were designed for ORF5 gene fragment of PRRSV by reverse transcription-loop mediated isothermal amplification (RT-LAMP). The Bst DNA polymerase was amplified by reverse transcriptase at 65 鈩，
本文编号：2157608
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