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Detection of nucleic acid of classical swine fever virus by reverse transcription loop-mediated isothermal amplification (RT-LAMP)

机译:通过逆转录环介导的等温扩增(RT-LAMP)检测经典猪瘟病毒的核酸

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Classical swine fever virus (CSFV) is the causative agent of Classical swine fever which is a highly contagious disease affecting swine and resulting in severe economic losses. In this study, we developed reverse transcription loopmediated isothermal amplification (RT-LAMP) assay targeting the 5’UTR gene for the detection of CSFV. This amplification method can be obtained in 1 h under isothermal conditions (65°C) employing a set of six specific primers mixtures. Amplification product was visualized by using hydroxynaphthol blue (HNB) dye and agarose gel electrophoresis. The sensitivity was 100 copy numbers. No cross-reactivity related to Japanese encephalitis virus (JEV) and porcine reproductive and respiratory syndrome virus (PRRSV) was demonstrated. The results demonstrated that the RT-LAMP assay is a useful tool for the rapid and sensitive for CSFV detection in swine.
机译:古典猪瘟病毒(CSFV)是古典猪瘟的病原体,它是一种高度传染性疾病,会影响猪群并造成严重的经济损失。在这项研究中,我们开发了针对5’UTR基因的逆转录环介导的等温扩增(RT-LAMP)分析法,用于检测CSFV。该扩增方法可以在等温条件下(65°C)在1小时内使用一组六种特异性引物混合物获得。通过使用羟萘酚蓝(HNB)染料和琼脂糖凝胶电泳观察扩增产物。灵敏度为100拷贝数。没有发现与日本脑炎病毒(JEV)和猪繁殖与呼吸综合征病毒(PRRSV)有关的交叉反应。结果表明,RT-LAMP测定法是快速,灵敏地检测猪中CSFV的有用工具。

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