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水仙退化病毒(Narcissus degeneration virus,NDV)是水仙上发生较为严重的病毒之一。本文根据NDV已报道的外壳蛋白基因序列,设计特异性引物,以感病水仙的总RNA为模板,建立了快速检测NDV的巢式RT-PCR方法。结果表明,巢式RT-PCR具有良好的特异性和灵敏度,能够从感染NDV的水仙样品上扩增出与预期大小相符的特异性目的条带,同时与其他病毒无交叉反应;灵敏度测定结果表示,巢式RT-PCR灵敏度较高,是普通RT-PCR的104倍,当RNA稀释到109倍时仍能被检测到。本文建立的巢式RT-PCR为水仙上NDV的快速检测提供了技术参考依据。
Narcissus degeneration virus (NDV) is one of the more serious viruses on narcissus. In this paper, based on the coat protein gene sequence reported by NDV, specific primers were designed and the nested RT-PCR method for rapid detection of NDV was established by using the total RNA of susceptible Narcissus as a template. The results showed that nested RT-PCR had good specificity and sensitivity, and could amplify the specific bands corresponding to the expected size from the NDV-infected narcissus samples without cross-reactivity with other viruses. The results of sensitivity analysis , Nested RT-PCR high sensitivity, is 104 times that of ordinary RT-PCR, when the RNA is diluted to 109 times can still be detected. The established nested RT-PCR provides a technical reference for the rapid detection of NDV on Narcissus.