PDF(Pubmed)
Abstract：
Gold nanoparticle (AuNP) fabrication via the oxidation of D-glucose is applied for detecting two foodborne pathogens, Enterococcus faecium (E. faecium) and Staphylococcus aureus (S. aureus). D-glucose is used as a reducing agent due to its oxidation to gluconic acid by sodium hydroxide (NaOH), resulting in the formation of AuNPs. Based on this mechanism, we develop AuNP-based colorimetric detection in conjunction with loop-mediated isothermal amplification (LAMP) for accurately identifying the infectious bacteria. Here, Au+ ions bind to the base of double-stranded DNA. In the presence of D-glucose and NaOH, the LAMP amplicon-Au+ complex maintains its bound state at 65 °C for 10 min while it is reduced to AuNPs in a dispersed form, exhibiting a red color. We aimed to pre-mix D-glucose with LAMP reagents before amplification and induce successful colorimetry without inhibiting amplification to simplify the experimental process and decrease the reaction time. Therefore, the entire process, including LAMP and colorimetric detection, is accomplished in approximately 1 h. The limit of detection of E. faecium and S. aureus is confirmed using the introduced method as 101 CFU/mL and 100 fg/μL, respectively. We expect that colorimetric detection using D-glucose-mediated AuNP synthesis offers an application for simple and immediate molecular diagnosis.
摘要：
通过D-葡萄糖的氧化制备金纳米颗粒(AuNP)用于检测两种食源性病原体,屎肠球菌(E.粪便）和金黄色葡萄球菌（S.金黄色葡萄球菌）。D-葡萄糖由于其被氢氧化钠（NaOH）氧化为葡萄糖酸而用作还原剂，形成AuNPs。基于这一机制,我们开发了基于AuNP的比色检测与环介导等温扩增（LAMP）相结合，以准确识别感染性细菌。这里,Au+离子与双链DNA的碱基结合。在D-葡萄糖和NaOH的存在下，LAMP扩增子-Au+复合物在65°C下保持其结合状态10分钟，同时以分散形式还原为AuNP，展示红色。我们旨在在扩增前将D-葡萄糖与LAMP试剂预混合，并在不抑制扩增的情况下诱导成功的比色法，以简化实验过程并减少反应时间。因此,整个过程,包括LAMP和比色检测，在大约1小时内完成。使用引入的方法确认屎肠球菌和金黄色葡萄球菌的检测限为101CFU/mL和100fg/μL，分别。我们期望使用D-葡萄糖介导的AuNP合成的比色检测为简单和立即的分子诊断提供了应用。
