肉制品中鸡肉源成分的Proofman-LMTIA方法检测

王博锐1; 2; 许丹丹1; 2; 谷蒙林2; 3; 姚蔚1; 2; 王耀1; 肖付刚2; 王德国2*

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DOI:10.12161/j.issn.1005-6521.2023.16.023

食品研究与开发:2023,44(16):162-166

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肉制品中鸡肉源成分的Proofman-LMTIA方法检测

王博锐1，2，许丹丹1，2，谷蒙林2，3，姚蔚1，2，王耀1，肖付刚2，王德国2*^1,2,3

(1.河南科技大学食品与生物工程学院，河南洛阳 471000;2.许昌学院食品与药学院河南省食品安全生物标识快检技术重点实验室，河南许昌 461000;3.河南工业大学粮油食品学院，河南郑州 450001)

Proofman-LMTIA for Detection of Chicken-Derived Components in Meat Products

WANG Borui1，2，XU Dandan1，2，GU Menglin2，3，YAO Wei1，2，WANG Yao1，XIAO Fugang2，WANG Deguo2*^1,2,3

(1.College of Food and Bioengineering，Henan University of Science and Technology，Luoyang 471000，Henan，China;2.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;3.College of Grain and Food，Henan University of Technology，Zhengzhou 450001，Henan，China)

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投稿时间：2022-12-15

中文摘要: 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。

中文关键词: Proofman 探针梯型熔解温度等温扩增技术肉制品鸡肉快速检测

Abstract: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.

keywords: proofreading enzyme-mediated probe cleavage ladder-shape melting temperature isothermal amplification meat product chicken rapid detection

文章编号：202316023 中图分类号： 文献标志码：

基金项目: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）

作者	单位
王博锐1，2，许丹丹1，2，谷蒙林2，3，姚蔚1，2，王耀1，肖付刚2，王德国2*	1.河南科技大学食品与生物工程学院，河南洛阳 471000；2.许昌学院食品与药学院河南省食品安全生物标识快检技术重点实验室，河南许昌 461000；3.河南工业大学粮油食品学院，河南郑州 450001

Author Name	Affiliation
WANG Borui1，2，XU Dandan1，2，GU Menglin2，3，YAO Wei1，2，WANG Yao1，XIAO Fugang2，WANG Deguo2*	1.College of Food and Bioengineering，Henan University of Science and Technology，Luoyang 471000，Henan，China；2.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；3.College of Grain and Food，Henan University of Technology，Zhengzhou 450001，Henan，China

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