Mesophilic Argonaute-Mediated Polydisperse Droplet Biosensor for Amplification-Free, One-Pot, and Multiplexed Nucleic Acid Detection Using Deep Learning.
Anal Chem
; 96(5): 2068-2077, 2024 02 06.
Article
in En
| MEDLINE
| ID: mdl-38259216
ABSTRACT
Detection of nucleic acids from a single multiplexed and amplification-free test is critical for ensuring food safety, clinical diagnostics, and environmental monitoring. In this study, we introduced a mesophilic Argonaute protein from Clostridium butyricum (CbAgo), which exhibits nucleic acid endonuclease activity, to achieve a programmable, amplification-free system (PASS) for rapid nucleic acid quantification at ambient temperatures in one pot. By using CbAgo-mediated binding with specific guide DNA (gDNA) and subsequent targeted cleavage of wild-type target DNAs complementary to gDNA, PASS can detect multiple foodborne pathogen DNA (<102 CFU/mL) simultaneously. The fluorescence signals were then transferred to polydisperse emulsions and analyzed by using deep learning. This simplifies the process and increases the suitability of polydisperse emulsions compared to traditional digital PCR, which requires homogeneous droplets for accurate detection. We believe that PASS has the potential to become a next-generation point-of-care digital nucleic acid detection method.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Nucleic Acids
/
Biosensing Techniques
/
Deep Learning
Type of study:
Diagnostic_studies
Language:
En
Journal:
Anal Chem
Year:
2024
Document type:
Article
Affiliation country:
China
Country of publication:
United States