建立蜂蜜中痕量玉米糖浆 DNA 的快速提取和灵敏检测系统

IF 4.1 Q2 FOOD SCIENCE & TECHNOLOGY Food Chemistry Molecular Sciences Pub Date : 2024-04-24 DOI:10.1016/j.fochms.2024.100206
Huixing Ye , Wenqiang Chen , Tao Huang , Junfeng Xu , Xiaofu Wang
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引用次数: 0

摘要

蜂蜜中掺杂外源糖浆已成为一种普遍现象,而目前的检测技术需要大型仪器,既繁琐又耗时。本研究通过整合核酸快速提取(REMD)和重组酶聚合酶扩增(RPA)技术,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。首先,开发了一种快速提取方法,可在 5 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,建立了检测玉米内源基因(ZssIIb)的 RPA 方法,该方法可在 30 分钟内检测出 12 个拷贝/μL 的玉米内源基因,且不会与其他植物源基因发生交叉反应。这表明 RPA 技术具有很高的灵敏度和特异性。最后,利用 REMD-RPA 检测平台对不同浓度的玉米糖浆掺假进行了检测,30 分钟内可检测出 1 % 的掺假。为了验证该方法的有效性,对 22 个市售样品进行了测试,结果显示所建立的 RPA 能在 30 分钟内检测出 7 个掺假样品。总之,所开发的方法快速、灵敏、特异,为现场快速检测蜂蜜掺假提供了技术支持,可为开发其他现场检测方法提供参考。
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Establishment of rapid extraction and sensitive detection system of trace corn syrup DNA in honey

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 (ZssIIb) 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.

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来源期刊
Food Chemistry Molecular Sciences
Food Chemistry Molecular Sciences Agricultural and Biological Sciences-Food Science
CiteScore
6.00
自引率
0.00%
发文量
83
审稿时长
82 days
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