环介导等温扩增法快速检测组织样本中耐甲氧西林敏感和耐甲氧西林金黄色葡萄球菌的可行性。
Feasibility of Loop-Mediated Isothermal Amplification for Rapid Detection of Methicillin-Susceptible and Methicillin-Resistant in Tissue Samples.
机构信息
Department of Orthopaedic Surgery, National Medical Center, Seoul, Korea.
Department of Orthopaedic Surgery, Korea University Ansan Hospital, Ansan, Korea.
出版信息
Clin Orthop Surg. 2022 Sep;14(3):466-473. doi: 10.4055/cios21277. Epub 2022 Jul 21.
BACKGROUND
To date, few studies have investigated the feasibility of the loop-mediated isothermal amplification (LAMP) assay for identifying pathogens in tissue samples. This study aimed to investigate the feasibility of LAMP for the rapid detection of methicillin-susceptible or methicillin-resistant (MSSA or MRSA) in tissue samples, using a bead-beating DNA extraction method.
METHODS
Twenty tissue samples infected with either MSSA (n = 10) or MRSA (n = 10) were obtained from patients who underwent orthopedic surgery for suspected musculoskeletal infection between December 2019 and September 2020. DNA was extracted from the infected tissue samples using the bead-beating method. A multiplex LAMP assay was conducted to identify MSSA and MRSA infections. To recognize the genus, , and methicillin resistance, 3 sets of 6 primers for the 16S ribosomal ribonucleic acid (rRNA) and the and genes were used, respectively. The limit of detection and sensitivity (detection rate) of the LAMP assay for diagnosing MSSA and MRSA infection were analyzed.
RESULTS
The LAMP result was positive for samples containing 10 colony-forming unit (CFU)/mL for 16S rRNA, 10 CFU/mL for , and 10 CFU/mL for . The limits of detection for 16S rRNA and were not different between MSSA and MRSA. For the 10 MSSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and and a 100% negative reaction for . For the 10 MRSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and but only 90% positive reactions for . The sensitivity (detection rate) of the LAMP assay for identifying MSSA and MRSA in infected tissue samples was 100% and 90%, respectively.
CONCLUSIONS
The results of this study suggest that the LAMP assay performed with tissue DNA samples can be a useful diagnostic method for the rapid detection of musculoskeletal infections caused by MSSA and MRSA.
背景
迄今为止，鲜有研究调查环介导等温扩增（LAMP）检测法在组织样本中鉴定病原体的可行性。本研究旨在使用珠磨法 DNA 提取方法，调查 LAMP 快速检测组织样本中耐甲氧西林敏感或耐甲氧西林金黄色葡萄球菌（MSSA 或 MRSA）的可行性。
方法
本研究纳入 2019 年 12 月至 2020 年 9 月期间因疑似肌肉骨骼感染而行骨科手术的患者的 10 例 MSSA 感染和 10 例 MRSA 感染组织样本，采用珠磨法从感染组织样本中提取 DNA。采用多重 LAMP 检测法鉴定 MSSA 和 MRSA 感染。为识别 属和耐甲氧西林，分别使用针对 16S 核糖体核糖核酸（rRNA）和 及 基因的 3 组 6 个引物。分析 LAMP 检测法诊断 MSSA 和 MRSA 感染的检测限和灵敏度（检出率）。
结果
当 16S rRNA 、 及 基因的检测限分别为 10 个菌落形成单位（CFU）/mL、10 CFU/mL 和 10 CFU/mL 时，LAMP 结果为阳性。MSSA 和 MRSA 的 16S rRNA 和 检测限无差异。对于 10 例 MSSA 阳性样本，LAMP 检测法对 16S rRNA 和 均呈 100%阳性反应，对 呈 100%阴性反应。对于 10 例 MRSA 阳性样本，LAMP 检测法对 16S rRNA 和 均呈 100%阳性反应，对 仅呈 90%阳性反应。LAMP 检测法鉴定感染组织样本中 MSSA 和 MRSA 的灵敏度（检出率）分别为 100%和 90%。
结论
本研究结果表明，使用组织 DNA 样本进行 LAMP 检测法可能是一种快速检测 MSSA 和 MRSA 引起的肌肉骨骼感染的有用诊断方法。
Zotero 插件