梯度条带PCR快速检测CSBV方法的建立

A Novel Method for Identification of CSBV by Gradient Band PCR

通过与传统RT-PCR检测方法比较,建立了一种新的快速、准确鉴定中华蜜蜂囊状幼虫病病毒(CSBV)的方法,为中华蜜蜂囊状幼虫病的诊断与防治提供了重要的手段。对陕南、陕北、关中三地的健康蜂及CSBV感染蜂分别进行传统RT-PCR和梯度条带PCR两种方法的检测,通过琼脂糖凝胶电泳及测序结果对两种方法进行分析比较。结果发现传统RT-PCR和梯度条带PCR方法对陕西三地的感染病毒蜂鉴定结果一致,而梯度条带PCR方法相对更快速和直观。与传统RT-PCR鉴定方法比较,该新方法能够在电泳检测阶段通过DNA片段的梯度次序即确定是否存在病毒,能够省去PCR产物测序步骤,缩短了对该病毒的鉴定周期,对及时发现和鉴定病蜂、采取预防措施提供了重要的技术手段。因此,梯度条带PCR快速鉴定方法不仅为CSBV的检测提供了一种简单、快速、直观的方法,对类似动物病毒的快速鉴定也提供了很好的借鉴。

A novel method for identification of CSBV,gradient band PCR（GBP） was set up in this study.Compared with traditional RT-PCR,this new identification method was very quick and was easily manipulated,which provided a new method to diagnosis and control the Chinese honeybee sacbrood.The healthy samples and CSBV infected bee larvae from south of Shaanxi,north of Shaanxi and middle of Shaanxi were confirmed by both of traditional RT-PCR and gradient band PCR,respectively.The PCR products were observed by AGE and then sent for sequencing.The results showed that the identification accuracy were the same by both RT-PCR and GBP,and the GBP method was a very quick,convenient and comparatively easy approach that could identify the virus without the results of PCR product sequences.This new GBP method is not only an useful method to identify CSBV infected honeybees but also sheds a light on methodology of identification for other similar animal viruses.