Department of Molecular Sciences, Macquarie University, Sydney, Australia.
Analyst. 2020 Apr 7;145(7):2789-2794. doi: 10.1039/c9an01959f. Epub 2020 Feb 26.
Methicillin-resistant Staphylococcus aureus (MRSA) is considered to be one of the important hospital-acquired pathogens. MRSA is also commonly associated with hospital-acquired infections and mortality. Quantitative and precise detection of MRSA is essential for rapid diagnosis and subsequent effective disease management strategies. We herein developed a highly specific method for rapid MRSA detection that combines surface-enhanced Raman spectroscopy (SERS) nanotags and polymerase chain reaction (PCR). SERS provided the sensitivity and spectral multiplexing capability while PCR provided the specificity required for the assay. The method was tested by the simultaneous detection of two MRSA specific genes (mecA and femA) amplified from genomic DNA isolated from clinical specimens. Magnetic isolation and rapid duplex detection were performed to obtain a detectable signal down to 10 input copies within 80 min. This demonstrated the potential of the SERS-PCR based approach for the accurate identification of MRSA at an early-diagnosis stage. This study also provides an alternative approach to the existing methods for detecting clinical targets without compromising sensitivity and selectivity, and with minimal handling steps. We thus believe that this approach will find a broad application in clinical applications.
耐甲氧西林金黄色葡萄球菌（MRSA）被认为是重要的医院获得性病原体之一。MRSA 也常与医院获得性感染和死亡率相关。快速诊断和随后实施有效的疾病管理策略，都需要对 MRSA 进行定量和精确检测。本研究开发了一种结合表面增强拉曼光谱（SERS）纳米标签和聚合酶链反应（PCR）的快速、高特异性的 MRSA 检测方法。SERS 提供了灵敏度和光谱多重检测能力，而 PCR 则提供了检测所需的特异性。该方法通过同时检测从临床标本中分离的基因组 DNA 扩增的两个 MRSA 特异性基因（mecA 和 femA）进行测试。通过磁分离和快速双重检测，可在 80 分钟内获得低至 10 个输入拷贝的可检测信号。这表明基于 SERS-PCR 的方法在早期诊断阶段准确识别 MRSA 的潜力。本研究还提供了一种替代方法，可以在不影响灵敏度和选择性的情况下，对临床靶标进行检测，且处理步骤最少。因此，我们相信这种方法将在临床应用中得到广泛应用。
