Dual-mode visual detection strategies of viable pathogens for point-of-care testing
Biosensors and Bioelectronics
; 221, 2023.
Article
in English
| Scopus | ID: covidwho-2240295
ABSTRACT
Here, we introduce a power-free foldable poly(methyl methacrylate) (PMMA) microdevice fully integrating DNA extraction, amplification, and visual detection, realized in novel dual modes – colorimetric and aggregate formation – using 4-Aminoantipyrine (4-AP) for monitoring pathogens. The microdevice contains two parts reaction and detection zones. A sealing film was utilized to connect the two zones and make the device foldable. The FTA card was deposited in the reaction zone for DNA extraction, followed by loop-mediated isothermal amplification (LAMP) at 65 °C for 45 min. When the detection zone is folded toward the reaction zone, paper discs modified with 4-AP placed in the detection zone are delivered to the reaction zone. Specifically, in the presence of LAMP amplicons, 4-AP is oxidized into antipyrine red or generates aggregates by interacting with copper sulfate, forming copper hybrid nanostructure (Cu-hNs). In the absence of LAMP amplicons, 4-AP is not oxidized and maintains yellow color or fails to form aggregates. Furthermore, we introduced the ethidium homodimer-1 (EthD-1) to identify viable bacteria. EthD-1 penetrated the compromised membranes of nonviable cells and prevented further DNA amplification by intercalating with the DNA. In this way, only samples containing viable cells displayed color change or formed aggregates upon reaction with 4-AP. Using this method, SARS-CoV-2 RNA and Enterococcus faecium were identified by naked eye, with the limit of detection of 103 copies/μL and 102 CFU/mL, respectively, within 60 min. The introduced microdevice can be used for rapidly monitoring viable pathogens and controlling outbreaks of infectious disease in resource-limited settings. © 2022 Elsevier B.V.
Biosensing Techniques; COVID-19; Humans; Point-of-Care Testing; RNA, Viral; SARS-CoV-2; Aggregates; Color; Colorimetry; Copper compounds; DNA; Extraction; Nanostructures; Sulfur compounds; virus RNA; 4-aminoantipyrine; Copper hybrid nanostructure; Dual modes; Dual-mode visual detection; Ethidium homodimer; Ethidium homodimer-1; Foldable poly(methyl methacrylate) microdevice; Hybrid nanostructures; Micro-device; Poly(methyl methacrylate); Poly-methyl methacrylates; Viability assay; Visual detection; diagnosis; genetic procedures; genetics; human; point of care testing; Esters; Foldable PMMA microdevice
Full text:
Available
Collection:
Databases of international organizations
Database:
Scopus
Language:
English
Journal:
Biosensors and Bioelectronics
Year:
2023
Document Type:
Article
Similar
MEDLINE
...
LILACS
LIS