调味粉中罂粟碱LAMP检测方法建立及试剂盒研制

摘要

建立调味粉中罂粟碱LAMP(loop-mediated isothermal amplification of DNA,LAMP)检测方法,并研制出配套试剂盒.根据罂粟碱的特异性基因序列设计引物,对12种罂粟、11种香辛料和1种虞关人核酸样品进行特异性、最佳温度筛选及敏感性实验,根据优化后的反应条件,组装试剂盒,并采用试剂盒对市售的8种调味粉进行检测.结果表明:12种罂粟和虞美人发生了特异性反应,最佳反应温度为65℃,LAMP检测最低浓度达70 pg/μL,敏感性是PCR检测的10倍.市售8种调味粉中,有1种含有罂粟碱成分.实验所建立的检测方法及研制的试剂盒对调味粉中掺加罂粟粉检测具有重要意义.%The LAMP detection method for papaverine in seasoning powders is constructed and the corresponding kit is developed.The primer is designed based on the specific genes sequence of papaverine.The specificity test,screening test for optimum temperature and sensitivity test of 12 poppies,11 spices and nucleic acid sample of a corn poppy are made.The kit is assembled based on the optimized reaction conditions,and 8 commercially seasoning powders are detected by using kit.The results are determined as follows:12 poppies and corn poppy show the specific reaction.The optimum temperature is 65 ℃ and the minimum detectable concentration is 70 pg/μL,the sensitivity is 10 times larger than that of PCR.There is papaverine in one of the 8 commercially seasoning powders.The detection method and development of kit have great significance for monitoring the adulteration of poppy powder in seasoning powders.