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A Novel Linear Displacement Isothermal Amplification with Strand Displacement Probes (LDIA-SD) in a Pocket-size Device for Point-of-care Testing of Infectious Diseases

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Date 2023 Jan 2
PMID 36589905
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Abstract

Nucleic acid amplification is crucial for disease diagnosis, especially lethal infectious diseases such as COVID-19. Compared with PCR, isothermal amplification methods are advantageous for point-of-care testing (POCT). However, complicated primer design limits their application in detecting some short targets or sequences with abnormal GC content. Herein, we developed a novel linear displacement isothermal amplification (LDIA) method using two pairs of conventional primers and (Bst) DNA polymerase, and reactions could be accelerated by adding an extra primer. Pseudorabies virus (high GC content) and (low GC content) genes were used to evaluate the LDIA assay. Using strand displacement (SD) probes, a LDIA-SD method was developed to realize probe-based specific detection. Additionally, we incorporated a nucleic acid-free extraction step and a pocket-sized device to realize POCT applications of the LDIA-SD method. The LDIA-SD method has advantages including facile primer design, high sensitivity and specificity, and applicability for POCT, especially for amplification of complex sequences and detection of infectious diseases.

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References
1.
Phillips E, Moehling T, Bhadra S, Ellington A, Linnes J . Strand Displacement Probes Combined with Isothermal Nucleic Acid Amplification for Instrument-Free Detection from Complex Samples. Anal Chem. 2018; 90(11):6580-6586. PMC: 5990927. DOI: 10.1021/acs.analchem.8b00269. View

2.
Vincent M, Xu Y, Kong H . Helicase-dependent isothermal DNA amplification. EMBO Rep. 2004; 5(8):795-800. PMC: 1249482. DOI: 10.1038/sj.embor.7400200. View

3.
Dao Thi V, Herbst K, Boerner K, Meurer M, Kremer L, Kirrmaier D . A colorimetric RT-LAMP assay and LAMP-sequencing for detecting SARS-CoV-2 RNA in clinical samples. Sci Transl Med. 2020; 12(556). PMC: 7574920. DOI: 10.1126/scitranslmed.abc7075. View

4.
Gou H, Bian Z, Cai R, Jiang Z, Song S, Li Y . The Colorimetric Isothermal Multiple-Self-Matching-Initiated Amplification Using Cresol Red for Rapid and Sensitive Detection of Porcine Circovirus 3. Front Vet Sci. 2020; 7:407. PMC: 7417626. DOI: 10.3389/fvets.2020.00407. View

5.
Wen J, Gou H, Wang S, Lin Q, Chen K, Wu Y . Competitive activation cross amplification combined with smartphone-based quantification for point-of-care detection of single nucleotide polymorphism. Biosens Bioelectron. 2021; 183:113200. DOI: 10.1016/j.bios.2021.113200. View