Highly specific and sensitive detection of the Mycobacterium tuberculosis complex using multiplex loop-mediated isothermal amplification combined with a nanoparticle-based lateral flow biosensor
| Autor: | Yijiang Chen, Xingui Yang, Junfei Huang, Ziyu Xiao, Shijun Li, Huijuan Chen, Xu Chen, Wenlin Zheng, Wei Chen |
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| Rok vydání: | 2021 |
| Předmět: |
Tuberculosis
Clinical Microbiology - Research Paper Loop-mediated isothermal amplification Biosensing Techniques Sensitivity and Specificity Microbiology 03 medical and health sciences chemistry.chemical_compound Media Technology medicine Humans Multiplex 030304 developmental biology Reaction conditions 0303 health sciences biology 030306 microbiology Mycobacterium tuberculosis biology.organism_classification medicine.disease Molecular biology genomic DNA Molecular Diagnostic Techniques chemistry Mycobacterium tuberculosis complex Nanoparticles Nucleic Acid Amplification Techniques Biosensor DNA |
| Zdroj: | Braz J Microbiol |
| ISSN: | 1678-4405 1517-8382 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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