Ultrasensitive and Specific Clustered Regularly Interspaced Short Palindromic Repeats Empowered a Plasmonic Fiber Tip System for Amplification-Free Monkeypox Virus Detection and Genotyping

The urgent necessityfor highly sensitive diagnostic tools hasbeen accentuated by the ongoing mpox (monkeypox) virus pandemic dueto the complexity in identifying asymptomatic and presymptomatic carriers.Traditional polymerase chain reaction-based tests, despite their effectiveness,are hampered by limited specificity, expensive and bulky equipment,labor-intensive operations, and time-consuming procedures. In thisstudy, we present a clustered regularly interspaced short palindromicrepeats (CRISPR)/Cas12a-based diagnostic platform with a surface plasmonresonance-based fiber tip (CRISPR-SPR-FT) biosensor. The compact CRISPR-SPR-FTbiosensor, with a 125 & mu;m diameter, offers high stability andportability, enabling exceptional specificity for mpox diagnosis andprecise identification of samples with a fatal mutation site (L108F)in the F8L gene. The CRISPR-SPR-FT system can analyzeviral double-stranded DNA from mpox virus without amplification inunder 1.5 h with a limit of detection below 5 aM in plasmids and about59.5 copies/& mu;L when in pseudovirus-spiked blood samples. OurCRISPR-SPR-FT biosensor thus offers fast, sensitive, portable, andaccurate target nucleic acid sequence detection.

Automated summary

This study presents a CRISPR/Cas12a-based diagnostic platform with a surface plasmon resonance-based fiber tip (CRISPR-SPR-FT) biosensor for detecting the mpox virus. The compact CRISPR-SPR-FT biosensor offers high stability and portability, and can analyze mpox viral DNA without amplification in under 1.5 hours with a limit of detection below 5 aM.