Staphylococcus aureus (S. aureus), including methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA), is an eminent human pathogen that can colonize the human host and cause severe life-threatening infections. The development of a reliable, simple and rapid assay for detecting S. aureus and identifying MRSA is important for diagnosis and follow-up treatment. A novel molecular diagnosis technique, named multiplex loop-mediated isothermal amplification linked to a nanoparticle-based lateral flow biosensor (m-LAMP-LFB), was applied to detect all S. aureus species and identify MRSA. Two sets of primers were designed based on the femA gene (S. aureus-specific gene) and the mecA gene (encoding penicillin-binding protein 2a), and the multiple-LAMP products were analyzed using LFB. The m-LAMP-LFB amplification conditions, including the target DNA concentration, reaction temperature and time, were optimized. The sensitivity and specificity of the m-LAMP-LFB method were tested in the current study, and the multiple-LAMP-LFB technology was applied to detect the MSSA and MRSA strains from clinical samples. The S. aureus- and MRSA-specific primers based on the femA and mecA genes allowed the multiple-LAMP technology to detect S. aureus and MRSA, respectively. The multiple-LAMP conditions were optimized at 63 °C for 40 min. The full process, including genomic DNA template preparation, LAMP, and product identification, could be achieved in 80 min. The limit of detection (LoD) of the multiple-LAMP assay for femA and mecA detection was 100 fg of genomic DNA template per reaction. The specificity of m-LAMP-LFB detection was 100 %, and no cross-reactions to non-S. aureus strains were observed. The multiple-LAMP-LFB technique developed in the current study is a reliable, simple, rapid, specific and sensitive method to identify MSSA and MRSA infections for appropriate antibiotic therapy.

中文翻译：

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使用与基于纳米粒子的侧向流生物传感器相连的多重回路介导的等温扩增，快速，直观地检测甲氧西林敏感和耐甲氧西林的金黄色葡萄球菌。

金黄色葡萄球菌（S. aureus），包括对甲氧西林敏感的金黄色葡萄球菌（MSSA）和耐甲氧西林的金黄色葡萄球菌（MRSA），是一种重要的人类病原体，可以定居于人类宿主并引起严重的威胁生命的感染。可靠，简单，快速的检测金黄色葡萄球菌和鉴定MRSA的方法的开发对于诊断和后续治疗很重要。一种新颖的分子诊断技术，即与基于纳米粒子的侧向流生物传感器（m-LAMP-LFB）相连的名为多重环介导的等温扩增，被用于检测所有金黄色葡萄球菌物种并鉴定MRSA。基于femA基因（金黄色葡萄球菌特异性基因）和mecA基因（编码青霉素结合蛋白2a）设计了两组引物，并使用LFB分析了多个LAMP产物。m-LAMP-LFB扩增条件，优化了目标DNA浓度，反应温度和时间。在本研究中测试了m-LAMP-LFB方法的敏感性和特异性，并且将multi-LAMP-LFB技术应用于检测临床样品中的MSSA和MRSA菌株。基于femA和mecA基因的金黄色葡萄球菌和MRSA特异性引物使multi-LAMP技术分别检测金黄色葡萄球菌和MRSA。将多个LAMP条件在63°C下优化40分钟。整个过程，包括基因组DNA模板的制备，LAMP和产品鉴定，都可以在80分钟内完成。femA和mecA检测的多重LAMP检测的检测限（LoD）为每个反应100 fg基因组DNA模板。m-LAMP-LFB检测的特异性为100％，与非S的无交叉反应。观察到金黄色葡萄球菌。当前研究中开发的multiple-LAMP-LFB技术是一种可靠，简单，快速，特异且敏感的方法，可用于识别MSSA和MRSA感染以进行适当的抗生素治疗。