DNA免提取结合qPCR冻干试剂的肉制品掺假鉴定方法

doi:10.12441/spyswjs.20220629003

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DNA免提取结合qPCR冻干试剂的肉制品掺假鉴定方法
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                        10.12441/spyswjs.20220629003
                    
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作者单位:天津大学化工学院，天津 300072
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通讯作者:葛志强（1963—），男，博士，副教授，硕士研究生导师，主要从事药物分析及食品安全与检测研究。E-mail：gezhiq@tju.edu.cn
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A Method for Identification of Adulteration in Meat Products by DNA Extraction-Free qPCR Lyophilized Reagents

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School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China

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【目的】建立肉制品中牛、鸭源性成分的现场快速定量检测方法。【方法】将DNA免提取法与荧光定量PCR（qPCR）冻干试剂法相结合，采用DNA免提取法制备qPCR反应模板。通过冻干工艺制备出的预混qPCR试剂，利用便携式qPCR系统对冻干试剂进行扩增。【结果】DNA免提取法简化了样品的提取、纯化步骤，该过程可在10 min内完成。DNA免提取法结合qPCR冻干试剂法的检测限可达1 pg/μL，从采样到得出结果只需45 min。预混qPCR冻干试剂可在4 ℃下保存6个月、室温下保存4个月、37 ℃下保存14 d。【结论】利用便携式qPCR系统即可对qPCR冻干检测试剂进行扩增，从而实现肉制品掺假的快速鉴别。

Abstract:

[Objective] This study aims to achieve rapid quantitative detection of beef- and duck-derived ingredients in meat products. [Method] The DNA extraction-free method was combined with fluorescent quantitative PCR (qPCR) lyophilized reagents was developed. The DNA extraction-free method was employed to prepare the template for qPCR. The premixed qPCR reagents were prepared by lyophilization. A portable qPCR system was used for amplification with the lyophilized reagents. [Result] The DNA extraction-free method with simplified sample extraction and purification steps achieved the detection within 10 min. The method combining the DNA extraction-free DNA method and qPCR lyophilized reagents can produce results within 45 min, with the limit of detection being 1 pg/μL. The premixed qPCR lyophilized reagents can be stored at 4 ℃ for 6 months, at room temperature for 4 months, and at 37 ℃ for 14 d. [Conclusion] The qPCR lyophilized reagents are appropriate for a portable qPCR system and can thus achieve rapid identification of adulteration in meat products.

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张娴雅,葛志强. DNA免提取结合qPCR冻干试剂的肉制品掺假鉴定方法[J].食品与生物技术学报,2024,43(12):146-153.

ZHANG Xianya, GE Zhiqiang. A Method for Identification of Adulteration in Meat Products by DNA Extraction-Free qPCR Lyophilized Reagents[J]. Journal of Food Science and Biotechnology,2024,43(12):146-153.

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收稿日期:2022-06-29
最后修改日期:2022-08-24
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在线发布日期: 2025-04-11
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