Establishment and application of multiple cross displacement amplification coupled with lateral flow biosensor (MCDA-LFB) for visual and rapid detection of Candida glabrata.

BACKGROUND

(Candida glabrata C. glabrata) is classified as a high-priority pathogen by the World Health Organization (WHO). It causes mucosal and deep infections in immunocompromised individuals, yet its diagnosis currently relies on slow culture processes. This highlights the urgent need for rapid and accurate detection methods.

METHODS

Ten primers targeting different regions of the C. glabrata ITS2 gene were designed to create a multiple cross displacement amplification (MCDA) assay coupled with a nanoparticle lateral flow biosensor (LFB). Following optimisation of temperature and time, the multiple cross displacement amplification coupled with lateral flow biosensor (MCDA-LFB) system was employed to detect C. glabrata DNA in clinical specimens.

RESULTS

Under optimal conditions (63 °C, 40 min), the C. glabrata-MCDA-LFB assay achieved a limit of detection (LoD) of 10 fg/µL.The assay successfully detected all C. glabrata strains tested and demonstrated no cross-reactivity with non-C. glabrata isolates. Findings showed that the C. glabrata-MCDA-LFB assay promptly and successfully detected all 57 C. glabrata-positive samples among 240 clinical specimens identified by traditional culture methods. The entire process, including sample processing (20 min), the MCDA reaction (40 min) and result documentation (2 min), was completed within 62 min.

CONCLUSION

The C. glabrata-MCDA-LFB assay developed in this study is a rapid, simplified, sensitive and specific technique that is straightforward to use. It can be used to screen for or diagnose C. glabrata infections in clinical settings, particularly in regions with limited resources.