Ultrasensitive and Specific Clustered Regularly Interspaced Short Palindromic Repeats Empowered a Plasmonic Fiber Tip System for Amplification-Free Monkeypox Virus Detection and Genotyping.
Yuzhi ChenZhi ChenTianzhong LiMeng QiuJinghan ZhangYan WangWu YuanAaron Ho-Pui HoOmar Al-HartomyWageh SwelmAbdullah Godran Al-SehemiXin ShiJingfeng LiZhongjian XieLi XuejinQizhen ZhangPublished in: ACS nano (2023)
The urgent necessity for highly sensitive diagnostic tools has been accentuated by the ongoing mpox (monkeypox) virus pandemic due to 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 this study, we present a clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a-based diagnostic platform with a surface plasmon resonance-based fiber tip (CRISPR-SPR-FT) biosensor. The compact CRISPR-SPR-FT biosensor, with a 125 μm diameter, offers high stability and portability, enabling exceptional specificity for mpox diagnosis and precise identification of samples with a fatal mutation site (L108F) in the F8L gene. The CRISPR-SPR-FT system can analyze viral double-stranded DNA from mpox virus without amplification in under 1.5 h with a limit of detection below 5 aM in plasmids and about 59.5 copies/μL when in pseudovirus-spiked blood samples. Our CRISPR-SPR-FT biosensor thus offers fast, sensitive, portable, and accurate target nucleic acid sequence detection.
Keyphrases
- label free
- crispr cas
- genome editing
- nucleic acid
- genome wide
- dna methylation
- sars cov
- gold nanoparticles
- high throughput
- randomized controlled trial
- systematic review
- coronavirus disease
- quantum dots
- sensitive detection
- copy number
- escherichia coli
- gene expression
- circulating tumor
- low cost
- structural basis
- binding protein
- transcription factor
- real time pcr
- genome wide identification