二重 PCR 快速检测中蜂囊状幼虫病病毒与东方蜜蜂微孢子虫方法的建立

摘要

中蜂囊状幼虫病和东方蜜蜂微孢子虫病是中蜂中常见的病害，对中蜂生产造成巨大危害，病原的早期快速检测是病害防控的重要手段。建立1种基于RNA提取的，快速、准确鉴定中蜂囊状幼虫病病毒（Chinese sacbrood virus ，CSBV）和东方蜜蜂微孢子虫（Nosema ceranae ，Nc）2种病原的二重PCR检测法。该方法同时检测CSBV和 Nc的灵敏度可分别低至6与3个拷贝，且准确可靠；与两病原的单一PCR检测法灵敏度和精度一致，而二重PCR方法更加简便快速，为中蜂囊状幼虫病和东方蜜蜂微孢子虫病的早期快速诊断提供了重要的手段。%Chinese sacbrood disease caused by Chinese sacbrood virus (CSBV) and nosemosis caused by Nosema ceranae (Nc) in Chinese honey bee (A pis cerana cerana) are common ,and devastating to native beekeeping in China .Therefore ,a quick and pre‐cise diagnosis of both pathogens at early stages is highly helpful for disease control .A duplex PCR method for rapid and precise detecting CSBV and Nc simultaneously was established based on RNA extraction .The lowest limits of simultaneous detection are 6 copies of CSBV and 3 copies of Nc ,moreover ,this method is precise and dependable .The sensitivity and precision of this method are similar to that of conventional single PCR method .The duplex PCR can serve as a simple ,rapid ,and precise method for early diagnosis of CSBV and Nc .