鸽A群轮状病毒(group A rotavirus,RVA)是影响养鸽业健康发展的主要病原之一。为实现鸽RVA的高通量快速检测,根据目前国内外流行的鸽RVA VP6基因保守区域设计特异性引物与探针,建立了一种可以特异性检测鸽RVA的实时荧光RT-PCR方法,并对...鸽A群轮状病毒(group A rotavirus,RVA)是影响养鸽业健康发展的主要病原之一。为实现鸽RVA的高通量快速检测,根据目前国内外流行的鸽RVA VP6基因保守区域设计特异性引物与探针,建立了一种可以特异性检测鸽RVA的实时荧光RT-PCR方法,并对该方法的灵敏度、特异性和重复性进行了评估。结果显示,该方法灵敏度高,最低检测限为7.22×10^(2)copies/μL;特异性强,与鸽群其他常见病毒无交叉反应;重复性好,组内与组间重复变异系数均小于1.5%。利用该方法和普通RT-PCR方法对200份临床样品进行检测,发现荧光RT-PCR方法的病毒阳性检出率高于普通RT-PCR方法,二者符合率为94.00%。结果表明,本研究建立的实时荧光RT-PCR方法敏感、特异、准确,重复性好,可为开展鸽RVA的临床检测及流行病学调查提供技术支持。展开更多
The purified recombinant nuclocapsid protein of PRRSV was used to conjoined with latex, and the conjoined concentration,temperature and time were optimized, a detection method for serum antibodies against Porcine repr...The purified recombinant nuclocapsid protein of PRRSV was used to conjoined with latex, and the conjoined concentration,temperature and time were optimized, a detection method for serum antibodies against Porcine reproductive and respiratory syndrome virus was established. Some tests for the method’s sensitivity, specificity and stability were confirmed. About 200 serum samples were detected by using the method, and these samples were also detected by recombinant N protein based ELISA and IDEXX ELISA kit. The result showed that the agreement ratio of the recombinant N protein based LAT with other two methods arrived at 93%, 78%, respectively.展开更多
文摘鸽A群轮状病毒(group A rotavirus,RVA)是影响养鸽业健康发展的主要病原之一。为实现鸽RVA的高通量快速检测,根据目前国内外流行的鸽RVA VP6基因保守区域设计特异性引物与探针,建立了一种可以特异性检测鸽RVA的实时荧光RT-PCR方法,并对该方法的灵敏度、特异性和重复性进行了评估。结果显示,该方法灵敏度高,最低检测限为7.22×10^(2)copies/μL;特异性强,与鸽群其他常见病毒无交叉反应;重复性好,组内与组间重复变异系数均小于1.5%。利用该方法和普通RT-PCR方法对200份临床样品进行检测,发现荧光RT-PCR方法的病毒阳性检出率高于普通RT-PCR方法,二者符合率为94.00%。结果表明,本研究建立的实时荧光RT-PCR方法敏感、特异、准确,重复性好,可为开展鸽RVA的临床检测及流行病学调查提供技术支持。
文摘The purified recombinant nuclocapsid protein of PRRSV was used to conjoined with latex, and the conjoined concentration,temperature and time were optimized, a detection method for serum antibodies against Porcine reproductive and respiratory syndrome virus was established. Some tests for the method’s sensitivity, specificity and stability were confirmed. About 200 serum samples were detected by using the method, and these samples were also detected by recombinant N protein based ELISA and IDEXX ELISA kit. The result showed that the agreement ratio of the recombinant N protein based LAT with other two methods arrived at 93%, 78%, respectively.
