食品中鲍鱼过敏源基因成分的检测

Detection of allergen gene of abalone in food

目的本文建立食品中鲍鱼过敏源基因成分实时荧光PCR检测方法。方法采用蛋白酶K消化法提取鲍鱼肌肉组织中基因组DNA,针对鲍鱼管家基因16S r RNA基因设计特异性引物和探针,确定实时荧光PCR反应体系和反应条件,建立了鲍鱼过敏源基因成分实时荧光PCR快速检测方法。选用鱿鱼、海参等海产品进行特异性试验;采用添加方法制备灵敏度试验样品,分别制备了鲍鱼过敏源基因成分含量分别为100%、10%、1%、0.1%、0.01%、0.001%的样品。结果对非鲍鱼类食品进行检测,结果显示出良好的特异性;灵敏度试验表明,本文建立方法的最低检测下限为0.01%。结论本文建立了特异性好,灵敏度高的鲍鱼过敏源基因成分检测方法。

Objective To establish a real-time fluorescent PCR method for detection of abalone gene anaphylactogen in food. Methods Genomic DNA of abalone muscle tissue was extracted by proteinase K digestion method. Specific primers and the probe were designed based on 16S rRNA genes of abalone. After PCR reaction system and conditions were optimized, the rapid real-time PCR detection method of abalone anaphylactogen was developed. The seafood including squid, sea cucumber and so on, was tested for specificity. The samples that contained abalone-derived components of 100%,10%,1%,0.1%,0.01%,and 0.001%respectively were made by blend method for sensitivity test. Results It was of high specificity by testing non abalone products, and the sensitivity test showed that the lowest amount of detecting was 0.01%. Conclusion The detection method of allergic abalone gene anaphylactogen with high specificity and sensitivity was established in this research.