4种提取试剂对三七Y病毒RNA提取效果的比较

Comparison of the Efficiency of Four Extraction Methods for Panax Virus Y RNA

为比较不同核酸提取试剂对三七Y病毒(Panax virus Y,Pn VY)RNA提取效果的差异,本文以感染病毒病的三七叶片为研究对象,选取了3种商业化的RNA提取试剂盒和优化的CTAB总核酸提取法对提取Pn VY RNA的质量、操作时间、使用成本、RT-PCR扩增效率及DNA去除效果等方面进行了比较。结果表明:北京全式金生物技术有限公司的Trans Zol Plant试剂提取的总RNA质量最高,RT-PCR扩增效率较高,DNA去除完全,但成本也最高;宝生物工程(大连)有限公司的RNAiso Plus Total RNA试剂提取的总RNA质量、RT-PCR扩增效率和使用成本居中,无DNA残留;北京百泰克生物技术有限公司的TRIpure试剂提取的总RNA质量、RT-PCR扩增效率最低,成本居中,也无DNA残留,但耗时最短;而CTAB法提取的总核酸质量居中,RT-PCR扩增效率最高,成本最低,能同时提取到RNA及DNA,但耗时最长。对于复合感染DNA和RNA两种核酸类型病毒的样品来讲,CTAB法为最佳选择;对仅由RNA病毒侵染的材料而言,综合考虑后,Ta KaRa的RNAiso Plus Total RNA提取试剂是较为理想的选择。

The RNA extraction efficiencies of three commercial kits and a modified CTAB method for Panax virus Y （PnVY）-infected Panax notoginseng leaf samples were comparatively analyzed on the aspects of extraction quality,operation time,cost,RT-PCR amplification efficiency and DNA elimination efficiency.The results showed that TransZol Plant reagent （TransGen Biotech） was the most efficient for total RNA extraction and DNA elimination,high efficient for RT-PCR amplification,and most costly; RNAiso Plus Total RNA extraction reagent （Takara Biotechnology Co.,Ltd.,Dalian） was in medium efficiency for total RNA extraction and RT-PCR amplification,with medium cost and nice performance for DNA elimination;TRIpure reagent （Bioteke Corporation） had the lowest efficiencies for total RNA extraction and RT-PCR amplification and the shortest operation time,with medium cost and nice performance for DNA elimination;the CTAB method had a medium efficiency for total nucleic acid extraction and the highest RT-PCR amplification efficiency,and it could simultaneously extract DNA and RNA with the lowest cost,but taking the longest time.In summary,our modified CTAB method is optimal for extraction of viral nucleic acids from the samples co-infected with DNA and RNA viruses,and RNAiso Plus Total RNA reagent （Takara Biotechnology Co.,Ltd.,Dalian） is the choicest option for viral RNA extraction by considering all the aspects.