绵羊痘病毒、山羊痘病毒及羊口疮病毒多重PCR检测方法的建立和应用

发布时间：2018-05-24 20:57

本文选题：多重PCR + 绵羊痘病毒　；参考：《动物医学进展》2017年03期

【摘要】：为了建立鉴别绵羊痘病毒(SPPV)、山羊痘病毒(GTPV)和羊口疮病毒(ORFV)的多重PCR检测方法,针对GenBank中3种病毒的基因组序列,合成了3对引物,通过优化多重PCR反应条件,建立了鉴别检测3种病毒的多重PCR方法。特异性试验表明,应用该方法可分别扩增出3种病毒对应的目的片段,对大肠埃希菌、沙门菌、猪圆环病毒2型(PCV2)、猪细小病毒(PPV)、Vero细胞、正常羊组织的DNA和灭菌双蒸水均无扩增;敏感性试验表明,该方法最低检测量分别为30.46pg/μL的绵羊痘病毒、28.9pg/μL的山羊痘病毒和26.94pg/μL的羊口疮病毒基因组DNA;应用本方法对85份临床病料进行检测,结果与其他已建立的单项PCR检测方法结果一致,说明该方法可以用于临床上SPPV、GTPV和ORFV的鉴别诊断。
[Abstract]:In order to establish a multiplex PCR method for the identification of sheep poxvirus, goat pox virus and sheep mouth ulcer virus, three pairs of primers were synthesized according to the genomic sequences of three kinds of viruses in GenBank, and the conditions of multiple PCR reaction were optimized. A multiplex PCR method for the identification and detection of three viruses was established. The specific test showed that the corresponding target fragments of three kinds of viruses could be amplified by using this method. The DNA and sterilized water of normal sheep tissues were not amplified for Escherichia coli, Salmonella, porcine circovirus type 2 PCV2 and porcine parvovirus PPV-Vero cells. The sensitivity test showed that the genomic DNA of sheep pox virus (30.46pg/ 渭 L) and sheep mouth sore virus (26.94pg/ 渭 L) were detected by this method, which were 28. 9pg / 渭 L of sheep pox virus and 28. 9pg / 渭 L of 26.94pg/ 渭 L, respectively, and 85 clinical samples were detected by this method. The results were consistent with those of other established single PCR detection methods, which indicated that this method could be used in the differential diagnosis of ORFV and SPPV.
【作者单位】：西北农林科技大学动物医学院;西北农林科技大学动物科技学院;
【基金】：陕西省重点产业创新链项目(2016KTZDNY02-06) 陕西省农业科技创新与攻关项目(2016NY-092) 西北农林科技大学试验示范站(基地)科技成果推广项目(TGZX2015-32) 陕西省科技统筹创新工程计划项目(2016KTZDNY02-05)
【分类号】：S852.654

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