不同处理方法对细菌气溶胶样本基因组DNA回收率的影响

Effects of different treatments on genomic recovery from air aerosol samples

为分析不同处理方法对空气中细菌气溶胶样本基因组回收率的影响,本研究利用玻璃纤维滤膜采集空气样品中的微生物气溶胶后,针对采样膜的不同处理(钢珠敲打、研磨、MOBIO PowerWater试剂盒)进行DNA提取方法比较,结果表明利用MOBIO PowerWater DNA Isolation Kit对采样膜进行基因组DNA的提取能获得高纯度的DNA(OD_(260nm)/OD_(280nm)=1.82);利用荧光定量PCR技术对所回收基因组进行定量分析,验证该处理方法的样品DNA得率最高,达到15.81%。起始模板浓度的对数与对应的CT值成反比且具有极强的线性关系,其线性相关系数R^2为0.9985。本研究将对养殖场等环境空气中病原体气溶胶样本处理具有重要指导意义。

Different treatment methods have a significant impact on air aerosol samples’ genome recovery. In this study, we used glass fiber filters to collect air samples of microbial aerosols, and compare different treatments on microbial DNA extraction methods, by using real-time PCR technology for quantitative analysis of the recycling genome to determine the appropriate air microorganism collection and microbial genomes extraction of DNA. The results showed that using MOBIO PowerWaterR DNA Isolation Kit for the extraction of genomic DNA sample can obtain high purity DNA （OD260nm/OD280nm=1.82）, and by using real-time PCR technology for quantitative analysis of the recycling genome, the DNA yield was the highest （15.81%）. The results might have important implication for the pathogen in the ambient air of aerosol samples processing.