中国输血杂志2011，Vol.24Issue(12)：1023-1027,5.
高灵敏度等温扩增法可视化检测血液中的病毒核酸
Highly sensitive and visible detection of virus by loop-mediated isothermal amplification
梁超 1成思佳 2武海萍 1周国华2
作者信息
- 1. 中国药科大学生命科学与技术学院 江苏南京210009
- 2. 南京军区军事医学研究所
- 折叠
摘要
Abstract
Objective To develop a sensitive,rapid,and convenient method for detection of viral nucleic acid in blood samples,such as HBV. Method The primers were designed according to the conservative regions of HBV genome,and the loop-mediated isothermal amplification ( LAMP) was carried out to amplify the target sequences of HBV. SYBR Green I was used as the indicator for the PCR products. The assay would be scored as positive if a fluorescent signal was detected. Results We have successfully developed a method for visible detection of blood virus and invented a portable device that combined the amplification and detection in one platform. The sensitivity of our proposed method is 10 copies of HBV per tube. The amplification and detection process could be done within 1 hour. In the testing of 62 clinical specimens, which have been determined as HBV positive by quantitative PCR,all specimens were positive and an accuracy of 100 % was a-chieved. We also detected 6 positive specimens from 42 clinical samples which have been determined as HBV negative by quantitative PCR. Conclusion The established method and device are highly sensitive and can realize single-tube detection of the blood virus,providing a powerful tool for point-of-care testing in resource-limited circumstances.关键词
乙型肝炎病毒/血液安全/环介导等温扩增/可视化/核酸检测Key words
HBV virus/Blood safety/loop-mediated isothermal amplification, Nucleic acid testing (NAT)分类
医药卫生引用本文复制引用
梁超,成思佳,武海萍,周国华..高灵敏度等温扩增法可视化检测血液中的病毒核酸[J].中国输血杂志,2011,24(12):1023-1027,5.
