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目的建立我国农业部未颁发农业转基因生物安全证书的转基因苜蓿草J101品系特异性环介导等温扩增(loop–mediated isothermal amplification,LAMP)检测方法。方法根据转基因苜蓿草品系J101 3’端外源插入片段与苜蓿草基因组DNA之间的邻接区序列设计6条引物,建立转基因苜蓿草J101品系特异性LAMP检测方法,并对本方法的特异性、灵敏度进行了测定。结果建立的检测方法特异于转基因苜蓿草J101成分检测,检测最低DNA浓度为(1imit of detection,LOD)为16 pg,相当于10拷贝转基因苜蓿草J101基因组DNA。结论本研究建立的转基因苜蓿草J101品系特异性LAMP检测方法特异性好,灵敏度高,能够快速、准确、稳定地对转基因苜蓿草J101成分进行检测分析。
Objective To establish a loop-mediated isothermal amplification (LAMP) method for detecting the transgenic alfalfa J101 strain that has not been issued a certificate of agricultural genetically modified organisms by Ministry of Agriculture. Methods Six primers were designed according to the 3 ’exogenous insertion sequence of transgenic alfalfa line J101 and the genomic DNA of alfalfa to establish a specific LAMP detection method for the transgenic alfalfa J101 strain. The specificity, sensitivity The measurement was carried out. Results The detection method was specific to the detection of transgenic alfalfa J101. The minimum detectable DNA concentration (LOD) was 16 pg, which corresponded to 10 copies of transgenic alfalfa J101 genomic DNA. Conclusion The specific LAMP detection method of transgenic alfalfa J101 strain established in this study has good specificity and high sensitivity, which can detect and analyze the composition of transgenic alfalfa J101 rapidly, accurately and stably.