Rapid Detection of Measles Virus Using Reverse Transcriptase/Recombinase Polymerase Amplification Coupled with...

Area of Science:

• Virology
• Molecular Biology
• Biotechnology

Background:

• Measles virus is highly contagious, necessitating rapid and simplified diagnostic methods.
• Current diagnostic approaches can be time-consuming and require specialized laboratory equipment.
• Developing point-of-care tests (POCTs) is crucial for timely intervention and disease control.

Purpose of the Study:

• To develop a simple, visual, and rapid diagnostic assay for the detection of measles virus.
• To evaluate the diagnostic performance of the developed assay using clinical samples.
• To assess the potential of the assay for point-of-care applications.

Main Methods:

• Development of a reverse transcriptase/recombinase polymerase amplification assay coupled with CRISPR/Cas12a and immunochromatographic lateral flow detection (RT-RPA-CRISPR-LFD).
• Assay optimization for speed (under 1 hour) and temperature (42 °C).
• Evaluation of diagnostic sensitivity and specificity using RT-PCR-confirmed measles virus positive and negative saliva samples.

Main Results:

• The RT-RPA-CRISPR-LFD assay detected as low as 31 copies of measles virus RNA.
• The assay demonstrated high diagnostic accuracy with 96% sensitivity and 100% specificity.
• The entire testing process was completed in less than 1 hour, enabling rapid results.

Conclusions:

• The developed RT-RPA-CRISPR-LFD assay provides a simple, visual, and rapid method for measles virus detection.
• This assay holds significant potential for point-of-care diagnosis of measles, improving accessibility and speed.
• The high accuracy and efficiency of this method offer new perspectives for managing measles outbreaks.