Objective To study the accurate and effective method for the extraction of influenza a H1N1 influenza virus.Methods A total of 40 pharyngeal swab points from patients with influenza A (H1N1) from December 2014 to December 2015 in our hospital were selected as the research object.The obtained specimens were 10-1 ～ 10-4 series dilution,obtained with different concentrations of influenza virus samples preparation of inspection.This research mainly uses three kinds of reagent box to carry on the experiment.A was Maxwell16 Promega automatic nucleic acid extraction apparatus and the extractor kit,as group A;B was the Qiagen RNeasy Mini Kit;C was domestic H company RNA extraction kit.The nucleic acid extraction of the three methods was compared.Results The total RNA concentration of A group was the highest,followed by B group,in C group was the lowest.The concentration of extraction rate in Group A at each dilution were highest and the difference was statistically significant (P ＜ 0.05).With increase of concentration,the influenza virus extraction rate decreased gradually in Group B and group C,especially in group C,when diluted to 10-4,influenza virus couldnt be extracted.Conclusion The effect of Automatic nucleic acid extraction apparatus for nucleic acid extraction is better,even under high dilution conditions,it can also achieve high nucleic acid extraction rate.It is a rapid and effective detection method to avoid the error of the manual extraction,which is worthy of promotion and application in clinical.%目的 研究甲型H1N1流感病毒核酸提取的比较准确和有效的方法.方法 选用2014年12月至2015年12月就诊的甲型H1N1流感患者的咽拭子共40份作为研究对象.对所获得的标本进行10-1～10-4的系列稀释,获得不同浓度的流感病毒标本备检.主要采用三种试剂盒进行实验:一种是Promega公司的Maxwell16全自动核酸提取仪以及该提取仪的配套试剂盒,为A组,一种是QIAGEN公司的Rneasy Mini Kit,为B组,第三种则是国产的H生物公司的RNA提取试剂盒,为C组.对三种方法的核酸提取情况进行比较.结果 对同一份咽拭子的核酸进行提取,A组所提取的总RNA浓度最高,B组的总RNA浓度次之,C组的总RNA浓度最低.A组在各个稀释浓度的流感病毒提取率均较高,差异均具有统计学意义(P＜0.05);C组和B组随着稀释浓度的增大,流感病毒的提取率逐渐降低,尤其是C组当稀释浓度为10-4时,提取不到流感病毒.结论 全自动核酸提取仪对核酸的提取效果较好,即使在高倍稀释的情况下也能够达到较高的核酸提取率,避免了手工提取的误差,是一种快捷有效的检测方法,值得在临床上推广使用.
展开▼
