Mycobacterium tuberculosis detection and genotyping using molecular probe-based biosensor

Abstract

Nucleic acid analysis is one of the most promising approaches in modern diagnostics, however it usually requires expensive amplification equipment. In this study, we propose and approve a method for bacterial pathogens detection and genotyping using a molecular probe-based biosensor without amplification. The sensor consists of a molecular beacon probe as a signal reporter with a fluorophore and a quencher attached to it, and two DNA strands, which have fragments complementary to the reporter and to the analyzed nucleic acid (analyte). The M. tuberculosis HigA1 gene was detected using this sensor, and a point mutation associated with antibiotic resistance was discriminated. As an additional demonstration of the applicability of the method without amplification, E.Coli 16S rRNA was detected. Amplification-free sample detection has been further tested and achieved.