Equipment-free, salt-mediated immobilization of nucleic acids for nucleic acid lateral flow assays.
Sens Actuators B Chem
; 351: 130975, 2022 Jan 15.
Article
in En
| MEDLINE
| ID: mdl-36568876
ABSTRACT
As the world has been facing several deadly virus crises, including Zika virus disease, Ebola virus disease, severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and Coronavirus disease 2019 (COVID-19), lateral flow assays (LFAs), which require minimal equipment for point-of-care of viral infectious diseases, are garnering much attention. Accordingly, there is an increasing demand to reduce the time and cost required for manufacturing LFAs. The current study introduces an equipment-free method of salt-mediated immobilization of nucleic acids (SAIoNs) for LFAs. Compared to general DNA immobilization methods such as streptavidin-biotin, UV-irradiation, and heat treatment, our method does not require special equipment (e.g., centrifuge, UV-crosslinker, heating device); therefore, it can be applied in a resource-limited environment with reduced production costs. The immobilization process was streamlined and completed within 30 min. Our method improved the color intensity signal approximately 14 times compared to the method without using SAIoNs and exhibited reproducibility with the long-term storage stability. The proposed method can be used to detect practical targets (e.g., SARS-CoV-2) and facilitates highly sensitive and selective detection of target nucleic acids with multiplexing capability and without any cross-reactivity. This novel immobilization strategy provides a basis for easily and inexpensively developing nucleic acid LFAs combined with various types of nucleic acid amplification.
AuNPs, gold nanoparticles; BSA, bovine serum albumin; Biotin; Equipment-free; Immobilization; LAMP, loop-mediated isothermal amplification; LF, lateral flow; LFA, lateral flow assay; LFIA, lateral flow immuno-assay; LOD, limit of detection; Lateral flow assay; MERS, Middle East respiratory syndrome; Metal salt; NALFA, nucleic acid lateral flow assay; NC, nitrocellulose; NTC, no-template control; Nucleic acid lateral flow assay; POC, point-of-care; RPA, recombinase polymerase amplification; RT-qPCR, quantitative reverse transcription-polymerase chain reaction; SAIoNs, salt-mediated immobilization of nucleic acids; SARS, severe acute respiratory syndrome; Streptavidin
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
Sens Actuators B Chem
Year:
2022
Document type:
Article