猴痘病毒PCR检测方法的建立

摘要

The F3L gene sequence ofmonkeypox virus was synthesized and cloned into the plasmid to be used as a target. A pair of PCR primers were designed and synthesized based on the nucleotide sequences of MPV in GenBank. After optimization of the detection method%从GenBank上调取猴痘病毒的基因序列，经过分析，找出了猴痘病毒的特异性靶基因序列F3L片段，人工合成猴痘病毒MPV（AF380138）F3L基因片段（48048-48509bp）并插入质粒，作为病毒检测的模拟阳性模板。根据该序列设计并合成PCR引物，对模拟的阳性模板进行扩增，结果能扩增出与目的片段大小一致的条带。建立了PCR检测方法，经条件优化后，对鸡痘、禽痘、羊痘等14种相似病毒核酸进行PCR扩增，发现具有良好的特异性。PCR方法能扩增出0．3Pg的阳性模板，显示建立的PCR方法具有高效、快速、特异