PDF(Pubmed)
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 (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.
摘要：
蜂蜜掺假外源性糖浆已成为一种普遍现象，和需要大型仪器的当前检测技术是麻烦和耗时的。在这项研究中,通过整合核酸的快速提取（REMD）和重组酶聚合酶扩增（RPA），开发了一种简单有效的方法，被称为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，可在五分钟内快速提取玉米糖浆DNA，以满足PCR和RPA测定的要求。然后，建立了玉米内源基因的RPA检测方法(ZssIIb),可以在30分钟内检测到12个拷贝/μL的内源玉米基因，而不会与其他植物来源的基因发生交叉反应。这表明RPA技术表现出高灵敏度和特异性。最后,利用REMD-RPA检测平台对不同浓度的玉米糖浆掺假进行检测,在30分钟内可检测到1％的掺假。对22个市售样品进行了测试，以验证该方法的有效性，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。总的来说,所开发的方法是快速的，敏感,具体,为蜂蜜掺假的现场快速检测提供技术支撑,可为其他现场检测方法的开发提供参考。
