摘要
目的优化乙型流感暴发监测中分子鉴定的实验条件。方法分别以病毒RNA提取试剂盒手工提取和全自动核酸提取工作站提取江苏省暴发标本RNA,采用实时荧光定量RT-PCR检测看家基因核糖核酸酶P(RNaseP)和乙型流感核蛋白(B-NP)基因;分别以本实验室建立的RT-PCR扩增体系和进口商品化RT-PCR扩增体系,对乙型流感基质蛋白(B-M)基因进行检测。结果看家基因RNaseP能有效监测采样和RNA纯化的有效性;手工提取RNA的效率高于自动提取;进口商品化RT-PCR体系的扩增M基因效果较好。结论以手工法提取RNA,以real-timeRT-PCR检测B-NP基因和以进口商品化RT-PCR扩增体系检测B-M基因,能相对客观地反映病毒流行现状。
Objective To optimize the experimental condition for the molecular detection of virus in the outbreak of influenza B. Methods RNA from influenza outbreak samples in Jiangsu province was purified by Viral RNA Purified Kit as well as automatic nucleic acid purification workstation. Housekeeping gene RNase Protein （ RNaseP） gene and influenza B nucleoprotein（B-NP）gene were detected by Real-time RT-PCR. Influenza B martrix protein M （B-M） gene was detected by RT-PCR using amplification system established by both ours and the imported commercial kit. Results Housekeeping gene RNaseP gene could monitor the validity of sample collection and RNA purification. Manual RNA purification efficiency was higher than that of the automatic way. Imported commercial RT-PCR kit used to amplify B-M gene was better than that of the system established by our lab. Conclusion Purificating RNA manually, detecting B-NP gene by Real-time RT-PCR or detecting B-M gene by imported commercial RT-PCR kit could relatively reflect the epidemic of influenza B.
出处
《公共卫生与预防医学》
2006年第5期35-38,共4页
Journal of Public Health and Preventive Medicine
