生物医学

Vol.3 No.3 (July 2013)

环介导等温扩增技术快速检测呼吸道博卡病毒
Rapid Detection of Respiratory Bocavirus by Loop Mediated Isothermal Amplification

 

作者:

闫明哲 , 胡适可 , 汤行春 :湖北大学,生命科学学院,武汉

向贇 , 张家云 , 艾洪武 :武汉市儿童医院,检验科,武汉

 

关键词:

环介导等温扩增技术博卡病毒实时荧光定量PCR LAMP—Loop Mediated Isothermal Amplification Human Bocavirus Realtime Fluores-Cence Quantitative PCR

 

摘要:

目的:建立一种检测呼吸道博卡病毒的环介导等温扩增方法(LAMP)。方法:通过博卡病毒NP1基因特异性序列的6个位点设计4条LAMP引物,建立LAMP检测体系。并对60例疑感染者的咽试子,同时采用实时荧光定量PCR和LAMP方法进行博卡病毒核酸的检测。结果:通过凝胶电泳能观察到LAMP方法扩增引物的存在,且采用此体系的检测60份咽试子中8份阳性反应和实时荧光定量PCR检测结果一致。结论:博卡病毒核酸的LAMP检测方法快速、简单、特异、灵敏、成本低,对实验仪器设备要求不高,适用于基层医疗机构临床检测和现场快速诊断。

Objective: To Build a method (LAMP, Loop mediated isothermal amplification) for detecting Human Bocavirus. Methods: By designing 4 LAMP primers specified for 6 sites in NP1 gene sequence of Bocavirus, we built a system to detect Human Bocavirus. And we detected 60 oropharyngeal swab samples by using this system and realtime fluores-cence quantitative PCR to check whether this system works well at the same time. Results: We found that we can get the production through the LAMP amplification and the gel electrophoresis results of the 60 samples, in which we found 8 positive samples, are consensus between the realtime fluores-cence quantitative PCR and the LAMP system. Conclusion: LAMP, a kind of quick, simple, specific, sensitive and low cost method in detecting the nucleic acids of virus, is an ideal alternative for quick screening assay and clinical detection in medical institutions because of the experimental instruments and requirements are not high.




文章引用:

闫明哲 , 向贇 , 胡适可 , 张家云 , 汤行春 , 艾洪武 (2013) 环介导等温扩增技术快速检测呼吸道博卡病毒。 生物医学, 3, 17-22. doi: 10.12677/HJBM.2013.33004

 

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