Okumura Kayo, Miyamoto Yuji, Mitarai Satoshi, Ato Manabu
Research Center for Biosafety, Laboratory Animal and Pathogen Bank, National Institute of Infectious Diseases, Japan Institute for Health Security, Aoba-cho, Higashimurayama, Tokyo, Japan.
Leprosy Research Center, National Institute of Infectious Diseases, Japan Institute for Health Security, Aoba-cho, Higashimurayama, Tokyo, Japan.
Microbiol Spectr. 2025 Jul;13(7):e0290624. doi: 10.1128/spectrum.02906-24. Epub 2025 Jun 11.
is the etiological agent of cutaneous nontuberculous mycobacteriosis, inducing granuloma formation in human skin. Other mycobacterial species, including , , and , also cause cutaneous infections, with and subsp. showing close genetic relatedness to is a recognized causative agent of Buruli ulcer, and numerous molecular detection and diagnostic methods have been established. However, there is a lack of specific detection methods for . Currently, confirming infection requires 16S rRNA or housekeeping gene sequencing, which is labor-intensive and time-consuming. In this study, we developed a one-step colorimetric loop-mediated isothermal amplification assay for simple and specific detection of . The results were confirmed by visual inspection of color change in the reaction solution, eliminating the need for specialized equipment. Our proposed workflow can be completed within 60 minutes from sample preparation to detection with high sensitivity. To the best of our knowledge, the loop-mediated isothermal amplification assay developed in this study represents the only test capable of specifically detecting utilizing only the loop-mediated isothermal amplification reaction. Consequently, this loop-mediated isothermal amplification assay has the potential for implementation in routine diagnosis of .
and , which are frequently reported as etiological agents of cutaneous nontuberculous mycobacteria, exhibit similar characteristics in terms of symptomatology and patient demographics. However, there is currently no simple identification method to distinguish between these two species. The one-step loop-mediated isothermal amplification method developed in this study represents a rapid diagnostic approach that yields results in a short time frame and does not necessitate specialized precision instruments, as the results can be visually determined. Consequently, the loop-mediated isothermal amplification method established in this study is anticipated to contribute significantly to the advancement of diagnostic techniques for cutaneous nontuberculous mycobacteria.
是皮肤非结核分枝杆菌病的病原体，可在人体皮肤中诱导肉芽肿形成。其他分枝杆菌种类，包括、和，也会引起皮肤感染，其中和亚种与在基因上显示出密切的相关性。是布氏溃疡的公认病原体，并且已经建立了许多分子检测和诊断方法。然而，目前缺乏针对的特异性检测方法。目前，确认感染需要进行16S rRNA或管家基因测序，这既费力又耗时。在本研究中，我们开发了一种一步比色环介导等温扩增法，用于简单、特异性地检测。通过目视检查反应溶液中的颜色变化来确认结果，无需专门设备。我们提出的工作流程从样品制备到检测可在60分钟内完成，且灵敏度高。据我们所知，本研究中开发的环介导等温扩增法是唯一一种仅利用环介导等温扩增反应就能特异性检测的试验。因此，这种环介导等温扩增法有潜力应用于的常规诊断。
和经常被报道为皮肤非结核分枝杆菌的病原体，在症状学和患者人口统计学方面表现出相似的特征。然而，目前尚无简单的鉴定方法来区分这两个物种。本研究中开发的一步环介导等温扩增法是一种快速诊断方法，能在短时间内得出结果，且不需要专门的精密仪器，因为结果可通过目视确定。因此，本研究中建立的环介导等温扩增法有望对皮肤非结核分枝杆菌诊断技术的进步做出重大贡献。
