牛源性成分LAMP检测方法的建立

Establishment of the Loop-mediated Isothermal Amplification for the Detection of Bovine-derived Materials

根据牛cytB基因设计3对特异性引物,运用GenieⅡ等温扩增荧光检测系统,以环介导等温扩增(LAMP)荧光检测方法为基础,建立了牛源性成分LAMP检测方法,并对该方法进行了反应体系和条件的优化。试验结果表明:该方法操作简单、特异性好;灵敏度比普通PCR方法高100倍,达到5.408×10^(-2)μg/μL;反应时间短,最快的10分钟内即可完成反应。

In this article,three pairs of specific primers were designed according to the bovine-derived cytB gene. Then based on the loop-mediated isothermal amplification（LAMP）fluorescence detection,a LAMP assay for detecting bovine-derived materials was established by using Genie Ⅱreal-time fluorescence isothermal amplification detection system. Subsequently the optimizations of the reaction system and conditions were conducted. Results showed that the method was simple to operate and presented good specificity. Meanwhile,compared with the normal level of general PCR,its sensitivity was 100-fold higher,and its detection limit was found to reach 5.408×10-2 μg/μL. The method also showed short reaction period,by which the fastest total reaction time could be less than 10 minutes.