小反刍兽疫一步法RT-PCR检测方法的建立
Acta Agriculturae Boreali-occidentalis Sinica（2015）
Abstract
为储备小反刍兽疫疫情防控技术,根据GenBank公布的小反刍兽疫病毒N基因序列设计合成特异性引物,以小反刍兽疫弱毒疫苗为材料,建立小反刍兽疫病毒一步法PT-PCR检测方法,并对所建方法进行条件优化和性能评价.结果显示,建立的方法能有效扩增大小约447 bp目的基因片段.该一步法RT-PCR最佳模板质量浓度为6.76×10-3 g/L,最佳退火温度60℃.该一步法RT-PCR检测方法性能评价结果显示,其最小模板检出质量浓度为6.76×10 6 g/L,且方法具良好的特异性和临床应用性,可用于小反刍兽疫多种样本的检测.
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