I-motif sensor for the fluorometric detection of Monkeypox

Area of Science:

• Molecular Biology
• Biotechnology
• Diagnostic Assays

Background:

• DNA virus-generating disorders, such as Mpox, pose significant public health challenges.
• Accurate and efficient diagnostic methods are crucial for timely disease management and control.
• Current diagnostic approaches may require complex procedures or longer turnaround times.

Purpose of the Study:

• To develop a novel, rapid, and sensitive isothermal fluorometric diagnostic method for DNA virus disorders.
• To investigate the mechanism of signal generation based on pH-induced DNA structural changes.
• To enable simultaneous detection of multiple viral targets efficiently.

Main Methods:

• Development of a multiplex-isothermal DNA amplification method (multiplex-LAMP).
• Utilized retinoblastoma (Rb) linear single-stranded DNA (ssDNA) that transforms into i-motifs under acidic conditions.
• Employed thiazole orange (TO), a fluorometric probe, for sensitive detection of i-motif structures.

Main Results:

• The multiplex-LAMP reaction released protons, significantly lowering pH and inducing ssDNA to i-motif transformation.
• Thiazole orange selectively bound to i-motifs, resulting in enhanced signal-on fluorescence.
• The developed system achieved simultaneous detection of multiple targets with high sensitivity (1.47 pg/mL) and efficiency (30 minutes).

Conclusions:

• The multiplex-LAMP/i-motif-TO system provides a rapid, sensitive, and efficient diagnostic tool for DNA virus disorders.
• This method offers a promising approach for accurate assessment of viral infections like Mpox.
• The technology has the potential to improve disease surveillance and clinical diagnostics.