Department of BioNano Technology, Gachon University, 1342 Seongnam-daero, Sujeong-gu, Seongnam-si 13120, Republic of Korea.
Department of Orthopedic Surgery, College of Medicine, Catholic Kwandong University, Incheon 21431, Republic of Korea.
Biosensors (Basel). 2023 May 10;13(5):535. doi: 10.3390/bios13050535.
Osteoarthritis (OA) is the most common joint disease, which accompanies pain and inconvenience in daily life owing to degradation of cartilage and adjacent tissues. In this study, we propose a simple point-of-care testing (POCT) kit for the detection of the MTF1 OA biomarker to achieve on-site clinical diagnosis of OA. The kit contains an FTA card for patient sample treatments, a sample tube for loop-mediated isothermal amplification (LAMP), and a phenolphthalein-soaked swab for naked eye detection. The MTF1 gene was isolated from synovial fluids using an FTA card and amplified using the LAMP method at 65 °C for 35 min. A test part of the phenolphthalein-soaked swab was decolorized in the presence of the MTF1 gene due to the pH change after the LAMP, but the color remained pink in the absence of the MTF1 gene. The control part of the swab served as a reference color in relation to the test part. When real-time LAMP (RT-LAMP), gel electrophoresis, and colorimetric detection of the MTF1 gene were performed, the limit of detection (LOD) was confirmed at 10 fg/μL, and the overall processes were completed in 1 h. The detection of an OA biomarker in the form of POCT was reported for the first time in this study. The introduced method is expected to serve as a POCT platform directly applicable by clinicians for easy and rapid identification of OA.
骨关节炎（OA）是最常见的关节疾病，由于软骨和邻近组织的退化，它会伴随日常生活中的疼痛和不便。在本研究中，我们提出了一种简单的即时检测（POCT）试剂盒，用于检测 MTF1 OA 生物标志物，以实现 OA 的现场临床诊断。该试剂盒包含用于患者样本处理的 FTA 卡、用于环介导等温扩增（LAMP）的样品管和用于肉眼检测的酚酞浸渍棉签。使用 FTA 卡从滑液中分离 MTF1 基因，并在 65°C 下使用 LAMP 方法在 35 分钟内扩增。由于 LAMP 后 pH 值发生变化，酚酞浸渍棉签的测试部分在存在 MTF1 基因的情况下会褪色，但在不存在 MTF1 基因的情况下颜色保持粉红色。棉签的对照部分用作与测试部分相关的参考颜色。当对 MTF1 基因进行实时 LAMP（RT-LAMP）、凝胶电泳和比色检测时，确定检测限（LOD）为 10 fg/μL，整个过程在 1 小时内完成。本研究首次报道了 POCT 形式的 OA 生物标志物检测。所介绍的方法有望成为临床医生直接应用的 POCT 平台，用于轻松快速识别 OA。
