转基因番木瓜GM YK品系的LAMP检测

Detection of transgenic papaya line GM YK by LAMP

应用环介导等温扩增技术建立一种快速灵敏的转基因番木瓜GM YK品系检测方法。针对转基因番木瓜GM YK品系外源基因与内源基因结合处序列设计1组特异性引物,利用SYBR Green荧光PCR仪实时监测扩增过程,对反应体系进行优化。用优化后的反应体系检测LAMP方法的灵敏性、特异性,并对市售番木瓜进行检测。LAMP检测方法反应速度快,在60 min内即可完成扩增,检测结果易于观察,通过肉眼观察染色情况即可分辨,该方法灵敏度高,检测限可达到5.78 pg,并且具有良好的特异性。LAMP检测方法可快速、灵敏、特异的检测转基因番木瓜GM YK品系,并适用于基层和现场检测。

The study established a rapid and sensitive transgenic papaya line GM YK detection methods using Loop-mediated isothermal amplification（LAMP）. Three pairs of primers were designed according to the sequences between the host plant genome DNA and the integrated gene of GM YK. Real-time monitoring of the LAMP reaction was achieved by real-time fluorescent SYBR Green PCR,the sensitivity and specificity of LAMP were determined and spot test was performed under optimized amplification condition. LAMP detection in this study can be completed within 60 min,and the results are easy to oberve by using SYBR green I. The LAMP was specific to transgenic papaya line GM YK with a detection limit of 5.78 pg. Therefore,the rapid and simple assay is a potential useful technique for line GM YK detection in field condition.