Rapid detection of mpox virus using recombinase aided amplification assay

Area of Science:

• Virology
• Molecular Diagnostics
• Public Health

Background:

• The global mpox virus outbreak presents a significant public health challenge, exacerbated by difficulties in early diagnosis and potential co-occurrence with COVID-19.
• Current mpox virus diagnosis relies on real-time polymerase chain reaction (RT-PCR), which is time-consuming and requires specialized equipment, limiting its use in resource-limited areas.

Purpose of the Study:

• To develop a rapid, sensitive, and specific diagnostic assay for mpox virus detection.
• To create a method suitable for widespread use, particularly in underdeveloped regions facing mpox outbreaks.

Main Methods:

• Development of a recombinase-aided amplification (RAA) assay targeting conserved regions (A27L and F3L genes) of the mpox virus.
• Evaluation of assay sensitivity (10 copies/reaction for A27L, 10^2 copies/reaction for F3L) and specificity using simulated and clinical samples.
• Comparison of RAA assay performance against standard RT-PCR methods.

Main Results:

• The RAA assay detected mpox virus within 5-10 minutes with high sensitivity and specificity (100% for simulated samples).
• The assay demonstrated comparable detection power to RT-PCR in clinical blinded sample testing.
• No cross-reactivity was observed with other pathogens, indicating high target specificity.

Conclusions:

• The developed RAA assay offers a rapid, sensitive, and specific method for mpox virus detection.
• Its low operational difficulty and suitability for resource-limited settings make it a valuable tool for global mpox surveillance and control.
• This assay can significantly aid in managing mpox outbreaks, especially in areas with limited access to advanced diagnostic facilities.