食品科学 ›› 2024, Vol. 45 ›› Issue (1): 210-216.doi: 10.7506/spkx1002-6630-20230315-149

• 安全检测 • 上一篇    

4 种植物源性成分多重real-time PCR检测方法的建立及其在食用淀粉中的应用

范维,高晓月,董雨馨,刘虹宇,李贺楠,赵文涛,郭文萍   

  1. (中国肉类食品综合研究中心,北京食品科学研究院,北京 100068)
  • 发布日期:2024-02-05
  • 基金资助:
    北京市科技计划项目(Z221100007122010);“十三五”国家重点研发计划重点专项(2018YFC1603400); 北京市优秀人才培养资助青年骨干个人项目(2017754154700G101)

Establishment of a Multiplex Real-Time PCR Method for Detection of Four Plant-Derived Components and Its Application to Edible Starches

FAN Wei, GAO Xiaoyue, DONG Yuxin, LIU Hongyu, LI Henan, ZHAO Wentao, GUO Wenping   

  1. (China Meat Research Center, Beijing Academy of Food Sciences, Beijing 100068, China)
  • Published:2024-02-05

摘要: 建立一种可同时快速检测红薯、木薯、马铃薯、玉米源性成分的多重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以红薯g3pdh基因、木薯g3pdh基因、马铃薯UGPase基因、玉米zSSIIb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,开展方法学验证,并对不同掺入比例模拟样品和实际淀粉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。与15 种非目标源性均无交叉反应;对目标DNA的检测灵敏度可达到3×10-3 ng/μL,且具有良好的线性关系和扩增效率;对淀粉样品的检出限可达0.1%,对50 份实际样品进行检测,结果与参比方法一致,说明建立的多重real-time PCR法可用于食用淀粉种类掺假鉴别检测。

关键词: 多重实时聚合酶链式反应;食用淀粉;木薯;红薯;马铃薯;玉米

Abstract: A multiplex real-time polymerase chain reaction (PCR) method for simultaneous and rapid detection of components derived from sweet potato, cassava, potato, and corn was established. The g3pdhs gene of sweet potato and cassava, the UGPase gene of potato and the zSSIIb gene of corn were used as target genes to design specific primers and TaqMan probe, and the 18S rRNA gene was used as the internal reference gene. Methodological validation was performed, and the proposed method was applied to simulated samples with different mixing ratios and actual starch samples. The results showed that this method was high-throughput, sensitive and specific. No cross-reaction was found with 15 non-target sources. The detection sensitivity for template DNA was 3 × 10-3 ng/μL, and the proposed method had a good linear relationship and high amplification efficiency. The detection limit for edible starch samples was 0.1%. The results of the PCR method for 50 actual samples were consistent with those of the reference method, indicating that this method could be used for the detection of common adulterants in edible starches.

Key words: multiplex real-time polymerase chain reaction; edible starch; cassava; sweet potato; potato; corn

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