Department of Microbiology, Kanya Gurukula Campus, Gurukula Kangri (Deemed to be University), Haridwar 249407, India.
Bioprospection and Product Development Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow 226015, India.
J Microbiol Methods. 2024 Dec;227:107064. doi: 10.1016/j.mimet.2024.107064. Epub 2024 Oct 22.
Tuberculosis (TB) is still the most frequent cause of morbidity and mortality in the world caused by Mycobacterium tuberculosis (MTB). Due to slow diagnostic and treatment options, the disease is a major concern for public health and also increases the burden on the global economy. Rapid, sensitive, and cheaper TB diagnosis test is urgent to lower their rates by point of care testing (POCT). Therefore, molecular detection techniques like recombinase polymerase assay (RPA) and Loop-mediated isothermal amplification (LAMP) play a significant role in this regard as they work on the principle of isothermal nucleic acid amplification. RPA and LAMP bridge the research gap between the previous PCR-based detection tool and other reported isothermal tools for MTB. In this review, we endeavor to provide an overview of the assay that will be a novel approach toward a rapid amplification and visualization of DNA by the naked eye in natural light. RPA and LAMP can prove to be a highly specific pathogen detection technique in combination with lateral flow (LF) strips and SYBR Green I. Optimization of amplification conditions also made the assay ideally suited to the resource-limited field application at POCT. Additionally, RPA and LAMP have paved the way for meeting a key component of the POC diagnosis of TB like universal drug susceptibility testing. However, RPA is more suitable at the POC level than LPA as it requires a lower amplification temperature of around 37-42 °C and a simpler primer design.
结核病(TB)仍然是由结核分枝杆菌(MTB)引起的全球发病率和死亡率最高的疾病。由于诊断和治疗方案缓慢,该疾病对公共卫生构成重大关切,也增加了全球经济的负担。快速、敏感和更便宜的结核病诊断测试对于通过即时检测(POCT)降低其发病率非常重要。因此,重组酶聚合酶检测(RPA)和环介导等温扩增(LAMP)等分子检测技术在这方面发挥了重要作用,因为它们基于等温核酸扩增的原理。RPA 和 LAMP 弥补了以前基于 PCR 的检测工具与其他报道的 MTB 等温工具之间的研究差距。在这篇综述中,我们努力提供一个概述的检测,将是一个新的方法对快速扩增和可视化的 DNA 肉眼在自然光下。RPA 和 LAMP 可以证明是一种高度特异性的病原体检测技术,与侧流(LF)条带和 SYBR Green I 结合使用。扩增条件的优化也使该检测非常适合在 POCT 进行资源有限的现场应用。此外,RPA 和 LAMP 为满足 POCT 诊断结核病的一个关键组成部分,如普遍药物敏感性测试铺平了道路。然而,与 LPA 相比,RPA 更适合 POCT 级别,因为它需要较低的扩增温度约 37-42°C 和更简单的引物设计。