鸭圆环病毒与鸭Ⅰ型肝炎病毒二重PCR检测方法的建立

Detection of Duck Circovirus and Duck Hepatitis Virus 1 by Multiplex PCR

目的:建立一种同时检测鸭圆环病毒(DuCV)和鸭Ⅰ型肝炎病毒(DHV)病原体的二重PCR技术。方法:根据DuCV和DHV的基因文库,分别设计了2对与DuCV和DHV某段基因序列互补的引物,用这2对引物对同一样品中DuCV和DHV模板进行二重PCR扩增。结果与结论:用建立的方法均同时得到了2条特异性的大小与实验设计相符(DuCV:245 bp;DHV:569 bp)的二重PCR扩增带,而且对其他禽病病原的PCR扩增结果均为阴性,能同时检出56pg的DHV RNA模板和6 pg的DuCV DNA模板。

Objective： A multiplex PCR was optimized to simuhaneously detect two pathogenic of duck hepatitis virus I（DHV） and duck circovirus（DuCV）. Methods： Two sets of specific primers were designed according to the sequences of DHV and DuCV according to the nucleotide sequence in GenBank. A multiplex PCR was carried out with the two specific primers. Results ＆ Conclusion： All the samples which contained DHV cDNA and DuCV DNA, could be amplified by the multiplex PCR using these two sets primers, yielding two specific bands of DHV 569 bp and DuCV 245 bp. No specific band was amplified from other avian pathogenic virus and bacte- ria. It turned out that as little as 56 pg of DHV RNA and 6 pg of DuCV DNA was detected using gel electropho- resis in this multiplex PCR.