摘要
根据羊传染性脓疱病毒GeneBank基因序列,设计合成一对引物。通过对PCR反应条件的优化,敏感性及特异性试验,成功建立了用于检测羊传染性脓疱病毒DNA的PCR检测方法。该方法成功扩增出390 bp的目的基因片段,敏感性试验证明可以检测出320×10^(-2)ng/L的DNA。特异性试验羊痘病毒、山羊关节炎脑炎病毒均未扩增出相应片段。重复性试验对3份羊传染性脓疱病毒阳性病料进行检测,结果均为阳性。临床应用对临床送检的16份疑似羊传染性脓疱病毒感染病料用上述建立优化的PCR方法进行检测,结果显示,9份为阳性,阳性样品的PCR扩增产物克隆后进行序列分析显示均为羊传染性脓疱病毒。证明该方法具有良好的敏感性和特异性,可以用于临床诊断。
According to the gene sequence of GeneBank of contagious ecthyma virus （CEV）, a pair of primers were designed. The PCR test to detect CEV was successfully established. By the optimization of PCR reaction conditions, a 390 bp fragments was amplified. The sensitivity of the assay can be up to 320× 10^-5 ng/μL. This method is specific, it can distinguish contagious ecthyma vi- rus from sheep pox virus and caprine arthritis encephalitis virus. Repeatability test in three suspicious-positive diseases of sheep contagious ecthyma for testing, the results were positive. Clinical application of the 16 suspected contagious ecthyma disease samples which were infected, showed 9 positive results. After cloning and analyzing sequence of the positive samples＇ PCR amplification product, it showed that all infected with CEV. The method has good sensitivity, speci- ficity, and it can be used for clinical diagnosis.
出处
《现代畜牧兽医》
2016年第10期1-6,共6页
Modern Journal of Animal Husbandry and Veterinary Medicine
基金
河南省农业科学院自主创新基金
