Tuberculosis remains a serious global public health challenge and requires the development of rapid, sensitive, and specific diagnostic tools for effective treatment and disease control. Bioimaging reporters are promising diagnostic tools that exploit the unique biochemical properties of Mycobacterium tuberculosis for real-time detection of viable cells from clinical samples. Moreover, these methods offer significant advantages over the conventional methods currently used in practice, including reduced assay time, increased specificity, and the ability to discriminate viable cells from dead cells. In this review, we highlight reporters of a different nature that the enable direct detection of Mycobacterium tuberculosis, eliminating complex sample preparation. Such reporters could serve as powerful tools in fluorescence microscopy, provide alternative strategies for automated culture-based diagnostic systems, and offer new approaches for developing point-of-care methods and diagnostic devices suitable for clinical practice.

Tuberculosis remains one of the leading infectious diseases worldwide, which is why new approaches are being developed as a foundation for future diagnostic methods. One promising strategy relies on porphyrins as biomarkers for the detection of Mycobacterium tuberculosis using fluorescence spectroscopy. Previous attempts have been made to detect microorganisms based on their porphyrin production. However, insufficient endogenous porphyrin production limits the applicability of this approach for mycobacterial detection. As a proof-of-principle study, we investigated the feasibility of using exogenous precursors to enhance porphyrin accumulation with the aim of detecting mycobacterial growth by means of fluorescence spectroscopy. The introduction of these precursors resulted in a significantly higher fluorescence signal intensity, making it possible to detect Mycobacterium tuberculosis and identify potential contamination.

Trehalose probes seem to be a promising area of tuberculosis diagnosis, these probes are capable of selectively penetrating Mycobacterium tuberculosis. These probes generate a fluorescent signal, enabling detection of mycobacteria. To analyze the current state of knowledge and prospects of trehalose probes as a new approach for rapid detection of Mycobacterium tuberculosis, a systematic review of scientific literature was conducted. The main types of probes include fluorogenic probes, "fluorophore-quencher" based probes, and photoactivatable probes. Trehalose probes enable selective detection of mycobacteria due to specific trehalose uptake and incorporation into the cell wall, followed by fluorescence activation. These probes allow for the detection of mycobacteria in sputum samples without complex sample preparation or washing. The method allows differentiation of viable and non-viable cells and can also be applied for drug susceptibility testing.