番鸭细小病毒与鸭圆环病毒二重PCR方法的建立

摘要

根据基因库中鸭圆环病毒和番鸭细小病毒的基因序列，分别设计了两对特异性引物，通过对二重PCR扩增条件的优化，研究建立了可同时鉴别检测鸭圆环病毒和番鸭细小病毒的二重PCR方法。用该方法对同一样品中鸭圆环病毒和番鸭细小病毒的模板进行PCR扩增，结果均得到了与实验设计相符的351 bp(鸭圆环病毒)和474 bp(番鸭细小病毒)的扩增条带，而对鸭I型肝炎病毒、鹅细小病毒、鸭副黏病毒、鸭瘟病毒和禽流感病毒等病原体的检测全为阴性。敏感性测定结果表明：该二重PCR技术最低能检出100 fg的鸭圆环病毒和番鸭细小病毒DNA模板。研究建立的鸭圆环病毒和番鸭细小病毒的二重PCR方法，具有快速、敏感、特异、定量和重复性好等优点，可用于临床上鸭圆环病毒和番鸭细小病毒感染的检测。%　　A duplex transcription polymerase chain reaction assay (duplex PCR) was developed and optimized to simultaneously detect duck circovirus and Muscovy duck parvovirus in one reaction. Two pairs of specific primers were designed according to the conserved regions on the sequences of duck circovirus and Muscovy duck parvovirus in GenBank. It was shown that samples containg duck circovirus and Muscovy duck parvovirus could be amplified into the specific bands,351 bp for duck circovirus and 474 bp for Muscovy duck parvovirus by this duplex PCR,but no specific bands of the same sizes were amplified from duck hepatitis virus type I,gosling plague virus,duck paramyxovirus,duck plague virus and avian influenza virus. As low as 100 fg of duck circovirus and Muscovy duck parvovirus DNA could be detected. This duplex PCR assay is a quick,sensitive,and specific test for detection of duck circovirus and Muscovy duck parvovirus,and will be useful for the control of these viruses in ducks.