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蜂蜜中痕量玉米糖浆DNA的快速提取及灵敏检测体系的建立

Establishment of rapid extraction and sensitive detection system of trace corn syrup DNA in honey.

作者信息

Ye Huixing, Chen Wenqiang, Huang Tao, Xu Junfeng, Wang Xiaofu

机构信息

College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo, Zhejiang 315211, China.

State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Key Laboratory of Traceability for Agricultural Genetically Modified Organisms, Ministry of Agriculture and Rural Affairs, P.R.China, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.

出版信息

Food Chem (Oxf). 2024 Apr 24;8:100206. doi: 10.1016/j.fochms.2024.100206. eCollection 2024 Jul 30.

Abstract

Honey adulteration with exogenous syrup has become a common phenomenon, and current detection techniques that require large instruments are cumbersome and time-consuming. In this study, a simple and efficient method was developed by integrating the rapid extraction of nucleic acids (REMD) and recombinase polymerase amplification (RPA), known as REMD-RPA, for the rapid screening of syrup adulteration in honey. First, a rapid extraction method was developed to rapidly extract corn syrup DNA in five minutes to meet the requirements of PCR and RPA assays. Then, the RPA method for detecting endogenous maize genes was established, which could detect 12 copies/μL of the endogenous maize gene within 30 min without cross-reacting with other plant-derived genes. This indicated that the RPA technique exhibited high sensitivity and specificity. Finally, the REMD-RPA detection platform was used to detect different concentrations of corn syrup adulteration, and 1 % adulteration could be detected within 30 min. The 22 commercially available samples were tested to validate the efficacy of this method, and the established RPA was able to detect seven adulterated samples in less than 30 min. Overall, the developed method is rapid, sensitive, and specific, providing technical support for the rapid field detection of honey adulteration and can serve as a reference for developing other field test methods.

摘要

蜂蜜被外源糖浆掺假已成为一种普遍现象,而目前需要大型仪器的检测技术既繁琐又耗时。在本研究中,通过整合核酸快速提取(REMD)和重组酶聚合酶扩增(RPA)开发了一种简单高效的方法,即REMD-RPA,用于快速筛查蜂蜜中的糖浆掺假情况。首先,开发了一种快速提取方法,可在五分钟内快速提取玉米糖浆DNA,以满足PCR和RPA检测的要求。然后,建立了检测内源性玉米基因的RPA方法,该方法可在30分钟内检测到12拷贝/μL的内源性玉米基因,且不与其他植物来源的基因发生交叉反应。这表明RPA技术具有高灵敏度和特异性。最后,使用REMD-RPA检测平台检测不同浓度的玉米糖浆掺假情况,30分钟内可检测到1%的掺假情况。对22个市售样品进行了测试以验证该方法的有效性,所建立的RPA能够在不到30分钟的时间内检测到7个掺假样品。总体而言,所开发的方法快速、灵敏且特异,为蜂蜜掺假的快速现场检测提供了技术支持,并可为开发其他现场检测方法提供参考。

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