摘要
为建立一种梯型熔解温度等温扩增技术(ladder-shape melting temperature isothermal amplification,LMTIA)结合Proofman探针(proofreading enzyme-mediated probe cleavage)检测肉制品中的鸡肉,选取鸡细胞核中单拷贝且特异性强的prolactin receptor基因为靶基因,设计LMTIA引物和Proofman探针,优化Proofman-LMTIA反应体系,进行特异性、灵敏度和最低检测限测定。结果表明,Proofman-LMTIA方法特异性强,可在30 min内完成检测,检测灵敏度为1 ng/μL,对人工模拟的混合肉样的最低检测限为0.1%。该方法适用于基层单位对肉制品掺假进行快速检测。
A proofreading enzyme-mediated probe cleavage combined with ladder-shape melting temperature isothermal amplification(Proofman-LMTIA)method was established for the detection of chicken-derived components in meat products.The specific single-copy nuclear prolactin receptor gene of chicken was selected as the target gene for the design of LMTIA primers and Proofman.The LMTIA system was optimized,and then the specificity,sensitivity,and limit of detection(LOD)of the established method were evaluated.The results showed that the established method realized complete amplification within 30 min with high specificity and demonstrated the sensitivity of 1 ng/μL and the LOD of 0.1%for artificially adulterated meat samples.The Proofman-LMTIA method established in this study is suitable for primary organizations to quickly detect adulterants of meat products.
作者
王博锐
许丹丹
谷蒙林
姚蔚
王耀
肖付刚
王德国
WANG Borui;XU Dandan;GU Menglin;YAO Wei;WANG Yao;XIAO Fugang;WANG Deguo(College of Food and Bioengineering,Henan University of Science and Technology,Luoyang 471000,Henan,China;Key Laboratory of Biomarker Based Rapid-detection Technology for Food Safety of Henan Province,College of Food and Pharmacy,Xuchang University,Xuchang 461000,Henan,China;College of Grain and Food,Henan University of Technology,Zhengzhou 450001,Henan,China)
出处
《食品研究与开发》
CAS
北大核心
2023年第16期162-166,共5页
Food Research and Development
基金
国家自然科学基金项目(31701689、32172300)
河南省科技攻关项目(212102110214)
河南省高校重点研发项目(21B550006)。
关键词
Proofman探针
梯型熔解温度等温扩增技术
肉制品
鸡肉
快速检测
proofreading enzyme-mediated probe cleavage
ladder-shape melting temperature isothermal amplification
meat product
chicken
rapid detection
作者简介
王博锐(1997-),男(汉),硕士研究生,研究方向:食品质量与安全快速检测;通信作者:王德国(1975-),男(汉),教授,博士,研究方向:食品质量与安全快速检测。
