改良Trizol-SiO2法在咽拭子及疱疹液EV71-RNA提取中的应用

Applicatin of a Improved Trizol-SiO2 Method in Throat Swab and EV71-RNA Extraction of Herpes Fluid

目的 研究改良Trizol-SiO2法在咽拭子及疱疹液EV71-RNA提取中相对于目前常用的Trizol-氯仿-二氧化硅提取法的优势.方法 选取2010年6月-2013年1月传染科及发热门诊300例手足口病患儿咽拭子和（或）疱疹液拭子标本分别采用Trizol-氯仿-二氧化硅提取法和改良Trizol-SiO2法进行实验结果统计学分析.结果 采用Trizol-氯仿-二氧化硅提取法的232例EV71-RNA阳性标本,采用改良Trizol-SiO2法所获得的定性结果均为阳性;采用Trizol-氯仿-二氧化硅提取法的68例EV71-RNA阴性标本,采用改良Trizol-SiO2法所获得的定性结果均为阴性,两种方法差异无统计学意义（P=0.993）.同时所获的EV71-RNA在进行荧光定量PCR扩增后的扩增曲线Ct值,两种方法差异无统计学意义（P=0.874）.结论 与目前实验室常用的提取EV71-RNA Trizol-氯仿-二氧化硅法相比较,改良Trizol-SiO2法剔除了氯仿试剂并延长了SiO2吸附核酸物质的时间,此方法实现了EV71-RNA的快速提取,适用于大样本量的操作,具有很好的临床应用价值.

Objective To research and improve Trizol-SiO2 Act with respect to the commonly used Trizol-chloroform in the throat swab and herpes fluid EV71-RNA extraction-the advantages of silica extraction method.Methods Selected HEMD children＇ throat swab and （or） herpes fluid swabs of 300 cases in fever clinic and department of infectious diseases from June 2010 to January 2013 were analyzed using Trizol-chloroform-silicaextraction and improved Trizol-SiO2 Method.Results The results of positive samples of all 232 cases of EV71-RNA with Trizol-chloroform-silica,which measured using the modified Trizol-SiO2 were positive.The results of negative samples of all 68 cases of EV71-RNA with Trizol-chloroform-silica,which measured using the modified Trizol-SiO2 were negative.The two methods had no statistical difference （P=0.993）.Meanwhile the amplifed curve Ct value of EV71-RNA obtained by real-time PCR of two methods had no statistical difference （P=0.874）.Conclusion Compared with Trizol-chloroform-silica commonly used to extract EV71-RNA in laboratories,the improved Trizol-SiO2 eliminated the chloroform reagents and extended the time of SiO2 adsorpted nucleic acid material.With fast extraction of EV71-RNA,this method is suitable for large sample,and has a good clinical application value.