用于快速检测非洲猪瘟病毒的重组酶辅助扩增结合侧向流动试纸条检测方法的开发
Development of a Recombinase-aided Amplification Combined With Lateral Flow Dipstick Assay for the Rapid Detection of the African Swine Fever Virus.
作者信息
Li Jiang Shuai, Hao Yan Zhe, Hou Mei Ling, Zhang Xuan, Zhang Xiao Guang, Cao Yu Xi, Li Jin Ming, Ma Jing, Zhou Zhi Xiang
机构信息
Faculty of environment and life, Beijing University of Technology, Beijing 100024, China.
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
出版信息
Biomed Environ Sci. 2022 Feb 20;35(2):133-140. doi: 10.3967/bes2022.018.
OBJECTIVE
To establish a sensitive, simple and rapid detection method for African swine fever virus (ASFV) B646L gene.
METHODS
A recombinase-aided amplification-lateral flow dipstick (RAA-LFD) assay was developed in this study. Recombinase-aided amplification (RAA) is used to amplify template DNA, and lateral flow dipstick (LFD) is used to interpret the results after the amplification is completed. The lower limits of detection and specificity of the RAA assay were verified using recombinant plasmid and pathogenic nucleic acid. In addition, 30 clinical samples were tested to evaluate the performance of the RAA assay.
RESULTS
The RAA-LFD assay was completed within 15 min at 37 °C, including 10 min for nucleic acid amplification and 5 minutes for LFD reading results. The detection limit of this assay was found to be 200 copies per reaction. And there was no cross-reactivity with other swine viruses.
CONCLUSION
A highly sensitive, specific, and simple RAA-LFD method was developed for the rapid detection of the ASFV.
目的
建立一种针对非洲猪瘟病毒（ASFV）B646L基因的灵敏、简便、快速的检测方法。
方法
本研究开发了一种重组酶辅助扩增-侧向流动试纸条（RAA-LFD）检测方法。重组酶辅助扩增（RAA）用于扩增模板DNA，侧向流动试纸条（LFD）用于在扩增完成后解读结果。使用重组质粒和致病核酸验证了RAA检测方法的检测下限和特异性。此外，检测了30份临床样本以评估RAA检测方法的性能。
结果
RAA-LFD检测方法在37℃下15分钟内完成，包括10分钟的核酸扩增和5分钟的LFD读取结果。该检测方法的检测限为每个反应200个拷贝。并且与其他猪病毒无交叉反应。
结论
开发了一种用于快速检测ASFV的高灵敏度、特异性和简便的RAA-LFD方法。
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