Tuberculosis (TB) is the deadliest infectious caused by Mycobacterium tuberculosis complex (MTBC). Because most TB cases occur within low-income populations, developing a specific, sensitive, cost-saving, and rapid point-of-care test for the early diagnosis of TB is important for achieving the WHO’s End Tuberculosis Strategy. In the current study, a novel nucleic acid detection strategy that includes multiplex loop-mediated isothermal amplification combined with a nanoparticle-based lateral flow biosensor (mLAMP-LFB) was used to detect MTBC. The two sets of LAMP primers specific to the IS6110 and gyrB genes of MTBC were successfully designed and validated for the detection of MTBC. The preferred reaction conditions for this assay were confirmed to be 65 °C for 40 min, and the amplification products could be visually identified through LFB within 2 min. The full assay process, including genomic DNA template extraction, LAMP reaction, and product detection, could be completed in 80 min. The limit detection of the assay was 100 fg of DNA in pure culture. The specificity of the assay was 100%, and it had no cross-reactions to other strains. Thus, the m-LAMP-LFB technology established in the present study was an objective, rapid, simple, and sensitive assay for MTBC identification, which could be applied in a clinical setting, especially in resource-constrained regions of the world.

结核病 (TB) 是由结核分枝杆菌复合体 (MTBC)引起的最致命的传染病。由于大多数结核病病例发生在低收入人群中，因此为结核病的早期诊断开发一种特异性、敏感、节省成本和快速的床旁检测对于实现世卫组织的终结结核病战略非常重要。在目前的研究中，一种新的核酸检测策略包括多重环介导的等温扩增结合基于纳米颗粒的侧向流动生物传感器 (mLAMP-LFB) 用于检测 MTBC。IS6110和gyrB特异的两组LAMP引物成功设计并验证了MTBC基因用于检测MTBC。该测定的优选反应条件确定为65°C 40 min，扩增产物可在2 min内通过LFB目测识别。整个检测过程，包括基因组DNA模板提取、LAMP反应和产物检测，可在80分钟内完成。该测定的极限检测是纯培养物中的 100 fg DNA。该测定的特异性为100%，与其他菌株无交叉反应。因此，本研究中建立的 m-LAMP-LFB 技术是一种客观、快速、简单、灵敏的 MTBC 鉴定方法，可应用于临床环境，特别是在世界资源有限的地区。