摘要
目的探讨离心柱法与Trizol法在流感病毒核酸提取中的差异,以找到提高阳性检出率的方法。方法对592例临床流行感冒(流感)疑似患者采集咽拭子标本,采用离心柱法与Trizol法分别提取病毒核酸,采用巢式PCR进行扩增,扩增产物行1%琼脂糖凝胶电泳,以2 000 bp Marker为标准,在564 bp区域出现阳性条带者为流感阳性。结果 592例临床流感疑似患者咽拭子标本中,应用离心柱法提取核酸222例(37.5%)阳性,甲型流感患者212例,乙型流感患者10例。Trizol法提取核酸显示66例(11.1%)患者呈阳性,其中甲型和乙型流感患者分别为63例和3例。自2014年12月至2015年4月,北京地区流感病毒流行以甲型流感病毒为主,乙型流感病例较少。流感的发病与季节相关,集中暴发时期在12月份左右,高发感染人群的年龄段为21~40岁,与性别无关。在病毒提取效率上,离心柱法阳性率显著高于Trizol法,差异具有统计学意义(χ2=111.66,P<0.01)。结论北京地区流感病毒的流行以甲型流感为主。离心柱法提取核酸效率要高于Trizol法,有利于早期进行病原学诊断。
Objective To compare and evaluate the efficacy of the two different nucleic acid extraction methods for influenza determination. Methods There were 592 clinical suspected cases pharynx samples were collected. There were two methods were used to extract nucleic acid, such as column extraction and Trizol manual method. After extraction, RNA Nucleic acid were used in the RT-PCR and test with the electrophoresis. When there was a 564 bp marker positive, the sample would be sent to be tested. Results From December 2014 to April 2015, the influenza A was the main epidemic in Beijing; and less influenza B in Beijing. There were 66 （11.1%） Trizol positive while 222 （37.5%） positive for the column extraction. Conclusions The main epidemic influenza virus is the influenza A in Beijing. Column extraction is more efficient than the Trizol manual method. Use the column extraction, the diagnosis correction could be improved.
作者
刘亚楠
李珺
华文浩
宋淑静
马小亮
董建平
宋蕊
Liu Ya＇nan;Li Jun;Hua Wenhao;Song Shujing;Ma Xiaoliang;Dong Jianping;Song Rui(Department of Clinical Laboratory;Department of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China;Department of lnJectious Diseases, Beijing Haidian Hospital, Bejing Haidian Section of Peking University Third Hospital, Beijing 100083, China)
出处
《中华实验和临床感染病杂志（电子版）》
CAS
2017年第5期474-478,共5页
Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)
基金
首都医学发展科研专项项目(No.2014-4-2175)
