摘要
目的建立用于鹅细小病毒(GPV)和鹅副粘病毒(GPMV)快速鉴别的双重PCR检测方法。方法根据2种病原体的基因保守序列,分别设计与GPV的VP3和GPMV的F基因互补的2对引物,对混合样品中GPV的DNA及GPMV反转录后的cDNA模板进行双重PCR扩增。应用该方法对黑龙江省9市(县)GPV、GPMV病毒病的疑似病料进行检测。结果对cDNA模板进行双重PCR扩增得到与试验设计相符的574bp(GPV)和884bp(GPMV)2条特异性扩增带。双重PCR可以检测到1.32ng/L的GPMV和298ng/L的GPV核酸模板,DPV扩增结果为阴性。检测7个市(县)的发病鹅群均为鹅GPV感染,未检出GPMV。结论双重PCR方法是一种快速、敏感、特异的检测方法,可用于家禽GPV和GPMV感染。
Objective To establish double PCR for rapid detection of Goose parvovirus （GPV） and Goose paramyxorirus （GPMV）. Methods Two pairs of specific primers were designed according to the conservative sequence VP3 gene of GPV and F gene of GPMV in the GenBank. The DNA of GPV and cDNA of GPMV in artificial mixed samples were am- plified by the double PCR with the two pairs primers. The dead young goose from nine cities in Heilongjiang were defected with this method. Results The result showed that two specific bands of 574 bp （GPV） and 880 bp （GPMV） were consistent with tests designed. The sensitivity of detection for GPMV and GPV by double PCR reached approximately 1.32 ng/L and 298 ng/L. The samples of seven cities were GPV infection and GPMV was not defected. Conclusion The double PCR is a speedy, specific and sensitive method to detect GPV and GPMV.
出处
《中国病原生物学杂志》
CSCD
2008年第7期496-498,共3页
Journal of Pathogen Biology
基金
黑龙江农垦总局科技计划项目(HNKXIV-08-05-01)
大庆市科技攻关课题(SGG01-077)
