鸭呼肠孤病毒的RT-PCR检测方法的建立

Establishment of a RT-PCR method for duck reovirus detection

为快速检测临床鸭呼肠孤病毒的感染情况,根据鸭呼肠孤病毒σC蛋白的基因序列设计1对特异性PCR引物,并以呼肠孤病毒基因组为模板,建立了鸭呼肠孤病毒的特异性RT-PCR检测方法。采集江苏省多地疑似呼肠孤病毒感染病料,提取基因组DNA进行RT-PCR扩增,产物经测序鉴定后判断建立的方法对临床样品的检出率。结果显示,该方法可以特异性扩增鸭呼肠孤病毒σC基因保守区438 bp的序列,与其他鸭病毒无交叉反应,最低可检测1 fg基因组DNA,对于临床样品检出率为100%。该方法可以用于临床快速诊断鸭呼肠孤病毒感染。

To rapidly assess the clinical infection of duck reovirus, a pair of specific primers based on duck reovirusσC protein gene sequence was designed. Using the duck reovirus genome as a template, a specific RT？PCR method was es？tablished and applied to the samples collected from ducks infected by suspected reovirus in Jiangsu province. The RT？PCR technique established can specifically amplify the 438？bp sequence of reovirus σC conservative region, and detect the ge？nomic DNA of duck reovirus as low as 1 fg with detection rate of 100%. The RT？PCR can be used for rapid clinical diagno？sis of duck reovirus infection.