tetano
Editor, Senior Moderator
Anal Chem
. 2026 May 14.
doi: 10.1021/acs.analchem.5c07791. Online ahead of print.
CRISPR-Cas12a2-Based Multiplexed Diagnostic for Rapid and Highly Sensitive Detection of Respiratory Viruses
Chuncao Liu[SUP] 1 [/SUP], Meng Feng[SUP] 1 [/SUP], Helin Yu[SUP] 1 [/SUP], Xinlian Zhang[SUP] 2 [/SUP], Yan Li[SUP] 1 [/SUP], Guodong Sui[SUP] 3 [/SUP], Wenwen Jing[SUP] 1 [/SUP], Xunjia Cheng[SUP] 1 [/SUP]
Affiliations
Infectious diseases severely threaten global public health security, necessitating rapid and highly sensitive diagnosis. This study presents a novel multiplex diagnostic platform combining transcription-mediated amplification (TMA) with the CRISPR-Cas12a2 system for rapid and highly sensitive detection of respiratory viruses. The assay uses an integrated microfluidic chip, which can simultaneously identify influenza A/B and respiratory syncytial viruses (RSV-A/B) with optimized CRISPR RNAs and isothermal amplification, achieving detection limits as low as 10[SUP]2[/SUP] copies/μL within 60 min. The detection system showed excellent specificity; nonspecific reactions were not observed in the presence of nucleic acids from other respiratory pathogens. Clinical validation using nasopharyngeal swabs demonstrated high concordance with real-time quantitative reverse transcription polymerase chain reaction, with most positive samples detected within 40 min. The system eliminates DNA amplification steps, reduces contamination risk, and simplifies the workflow. Using two-step reactions on a centrifugal microfluidic chip, the TMA-CRISPR-Cas12a2 platform offers a promising integrated platform for multiplex respiratory pathogen screening, thereby supporting timely diagnosis and outbreak management.
. 2026 May 14.
doi: 10.1021/acs.analchem.5c07791. Online ahead of print.
CRISPR-Cas12a2-Based Multiplexed Diagnostic for Rapid and Highly Sensitive Detection of Respiratory Viruses
Chuncao Liu[SUP] 1 [/SUP], Meng Feng[SUP] 1 [/SUP], Helin Yu[SUP] 1 [/SUP], Xinlian Zhang[SUP] 2 [/SUP], Yan Li[SUP] 1 [/SUP], Guodong Sui[SUP] 3 [/SUP], Wenwen Jing[SUP] 1 [/SUP], Xunjia Cheng[SUP] 1 [/SUP]
Affiliations
- PMID: 42133534
- DOI: 10.1021/acs.analchem.5c07791
Infectious diseases severely threaten global public health security, necessitating rapid and highly sensitive diagnosis. This study presents a novel multiplex diagnostic platform combining transcription-mediated amplification (TMA) with the CRISPR-Cas12a2 system for rapid and highly sensitive detection of respiratory viruses. The assay uses an integrated microfluidic chip, which can simultaneously identify influenza A/B and respiratory syncytial viruses (RSV-A/B) with optimized CRISPR RNAs and isothermal amplification, achieving detection limits as low as 10[SUP]2[/SUP] copies/μL within 60 min. The detection system showed excellent specificity; nonspecific reactions were not observed in the presence of nucleic acids from other respiratory pathogens. Clinical validation using nasopharyngeal swabs demonstrated high concordance with real-time quantitative reverse transcription polymerase chain reaction, with most positive samples detected within 40 min. The system eliminates DNA amplification steps, reduces contamination risk, and simplifies the workflow. Using two-step reactions on a centrifugal microfluidic chip, the TMA-CRISPR-Cas12a2 platform offers a promising integrated platform for multiplex respiratory pathogen screening, thereby supporting timely diagnosis and outbreak management.