Single Universal Primer Recombinase Polymerase Amplification-based Lateral Flow Biosensor (SUP-RPA-LFB) for Multiplex Detection of Genetically Modified Maize,Analytica Chimica Acta

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Single Universal Primer Recombinase Polymerase Amplification-based Lateral Flow Biosensor (SUP-RPA-LFB) for Multiplex Detection of Genetically Modified Maize
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-08-01 , DOI: 10.1016/j.aca.2020.06.001
Kai Li ₁ , Yunbo Luo ₂ , Kunlun Huang ₂ , Zhansen Yang ₁ , Yusong Wan ₃ , Wentao Xu ₂

Affiliation

In this study, an isothermal paper biosensor, combining single universal primer recombinase polymerase amplification (SUP-RPA) and the lateral flow technique was developed for the multiplex detection of genetically modified maize (GMM). In pre-amplification stage, the event-specific primers contain a universal sequence at the 5' end, with a biotin-labeled deoxycytidine triphosphate (dCTP) deoxynucleotide providing additional amplification, which improves their amplification ability and ensures consistent multiplex amplification efficiency. In the signal recognition strategy, the SUP-RPA products are identified visually using the lateral flow biosensor (LFB) through dual hybridization. The accumulation of gold nanoparticles (AuNPs) produces a characteristic red band. Through this biosensor, a limit of detection of at least 50 copies was achieved, which is sensitive enough to detect MON810, MON863 and MON89034 simultaneously. The entire process of analysis was completed within 30 min and without any large-scale instrumentation. This biosensor, therefore, provides a novel rapid and portable multiple detection method for point-of-care applications, especially genetically modified organism (GMO) event-specific detection.

中文翻译：

基于单一通用引物重组酶聚合酶扩增的侧流生物传感器 (SUP-RPA-LFB) 用于多重检测转基因玉米

在这项研究中，开发了一种等温纸生物传感器，结合单一通用引物重组酶聚合酶扩增 (SUP-RPA) 和横向流动技术，用于转基因玉米 (GMM) 的多重检测。在预扩增阶段，事件特异性引物在 5' 端包含通用序列，生物素标记的脱氧胞苷三磷酸 (dCTP) 脱氧核苷酸提供额外的扩增，提高了它们的扩增能力并确保一致的多重扩增效率。在信号识别策略中，SUP-RPA 产品通过双杂交使用侧流生物传感器 (LFB) 进行视觉识别。金纳米粒子 (AuNPs) 的积累会产生特征性的红色带。通过这种生物传感器，达到了至少 50 个拷贝的检测限，其灵敏度足以同时检测 MON810、MON863 和 MON89034。整个分析过程在30分钟内完成，没有任何大型仪器。因此，该生物传感器为即时应用，尤其是转基因生物 (GMO) 事件特异性检测提供了一种新型的快速便携式多重检测方法。

更新日期：2020-08-01

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