Lin Chuangxin, Zeng Yongmei, Zhu Zhihong, Liao Jiayu, Yang Tiandan, Liu Yaqun, Wei Huagui, Li Jiamin, Ma Jibin, Wu Xiaoqing, Lin Guangyu, Lin Liyun, Chen Liying, Huang Huiying, Chen Weizhong, Wang Junli, Wen Feiqiu, Lin Min
Department of Pediatrics, Second Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong, China.
Department of Pediatric Hematology and Oncology, Shenzhen Children's Hospital, China Medical University, Shenzhen, Guangdong, China.
Microbiol Spectr. 2023 Mar 28;11(2):e0447622. doi: 10.1128/spectrum.04476-22.
Antimicrobial resistance (AMR) has posed a global threat to public health. The Staphylococcus aureus strains have especially developed AMR to practically all antimicrobial medications. There is an unmet need for rapid and accurate detection of the S. aureus AMR. In this study, we developed two versions of recombinase polymerase amplification (RPA), the fluorescent signal monitoring and lateral flow dipstick, for detecting the clinically relevant AMR genes retained by S. aureus isolates and simultaneously identifying such isolates at the species level. The sensitivity and specificity were validated with clinical samples. Our results showed that this RPA tool was able to detect antibiotic resistance for all the 54 collected S. aureus isolates with high sensitivity, specificity, and accuracy (all higher than 92%). Moreover, results of the RPA tool are 100% consistent with that of PCR. In sum, we successfully developed a rapid and accurate AMR diagnostic platform for S. aureus. The RPA might be used as an effective diagnostic test in clinical microbiology laboratories to improve the design and application of antibiotic therapy. Staphylococcus aureus is a species of Staphylococcus and belongs to Gram-positive. Meanwhile, S. aureus remains one of the most common nosocomial and community-acquired infections, causing blood flow, skin, soft tissue, and lower respiratory tract infections. The identification of the particular gene and the other eight genes of drug-resistant S. aureus can reliably and quickly diagnose the illness, allowing doctors to prescribe treatment regimens sooner. The detection target in this work is a particular gene of S. aureus, and a POCT is built to simultaneously recognize S. aureus and analyze genes representing four common antibiotic families. We developed and assessed a rapid and on-site diagnostic platform for the specific and sensitive detection of S. aureus. This method allows the determination of S. aureus infection and 10 different AMR genes representing four different families of antibiotics within 40 min. It was easily adaptable in low-resource circumstances and professional-lacking circumstances. It should be supported in overcoming the continuous difficulty of drug-resistant S. aureus infections, which is a shortage of diagnostic tools that can swiftly detect infectious bacteria and numerous antibiotic resistance indicators.
抗菌药物耐药性(AMR)已对全球公共卫生构成威胁。金黄色葡萄球菌菌株尤其对几乎所有抗菌药物都产生了耐药性。对金黄色葡萄球菌的AMR进行快速准确检测的需求尚未得到满足。在本研究中,我们开发了两种重组酶聚合酶扩增(RPA)方法,即荧光信号监测法和侧向流动试纸条法,用于检测金黄色葡萄球菌分离株中保留的临床相关AMR基因,并同时在种水平上鉴定此类分离株。通过临床样本验证了其敏感性和特异性。我们的结果表明,这种RPA工具能够以高敏感性、特异性和准确性(均高于92%)检测所有收集的54株金黄色葡萄球菌分离株的抗生素耐药性。此外,RPA工具的结果与PCR结果100%一致。总之,我们成功开发了一种用于金黄色葡萄球菌的快速准确的AMR诊断平台。RPA可作为临床微生物实验室的一种有效诊断测试,以改进抗生素治疗的设计和应用。金黄色葡萄球菌是葡萄球菌属的一种,属于革兰氏阳性菌。同时,金黄色葡萄球菌仍然是最常见的医院获得性和社区获得性感染之一,可引起血流、皮肤、软组织和下呼吸道感染。鉴定耐药金黄色葡萄球菌的特定基因和其他八个基因可以可靠且快速地诊断疾病,使医生能够更快地开出治疗方案。本研究的检测靶点是金黄色葡萄球菌的一个特定基因,并构建了一种即时检验(POCT)方法,以同时识别金黄色葡萄球菌并分析代表四个常见抗生素家族的基因。我们开发并评估了一种用于特异性和灵敏检测金黄色葡萄球菌的快速现场诊断平台。该方法能够在40分钟内确定金黄色葡萄球菌感染情况以及代表四个不同抗生素家族的10种不同AMR基因。它易于在资源匮乏和缺乏专业人员的情况下应用。在克服耐药金黄色葡萄球菌感染持续存在的困难方面,即缺乏能够快速检测感染细菌和多种抗生素耐药指标的诊断工具方面,它应得到支持。