The potential of tailed amplicons for SARS-CoV-2 detection in Nucleic Acid Lateral Flow Assays.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

SARS-CoV-2*/genetics ; SARS-CoV-2*/isolation & purification ; COVID-19*/diagnosis ; COVID-19*/virology ; RNA, Viral*/genetics; Humans ; Limit of Detection ; Nucleic Acid Amplification Techniques/methods ; Sensitivity and Specificity ; COVID-19 Nucleic Acid Testing/methods ; DNA, Single-Stranded/genetics ; DNA Primers/genetics ; DNA Probes

Nucleic Acid Lateral Flow Assays (NALFAs) are a promising solution for the point-of-care detection of viruses like SARS-CoV-2. However, they show some drawbacks, such as the great dependency on the use of antibodies and the need for post-amplification protocols that enable the preparation of amplicons for effective readings, as well as low sensitivity. Here, we developed amplicons of a specific SARS-CoV-2 gene tailed with single-strand DNA (ssDNA) sequences to hybridize with DNA probes immobilized on the NALFA strips, thus overcoming the aforementioned problems. Results have shown that tailed primers have not compromised the amplification efficiency and allowed the correct detection of the amplicons in the lateral flow strip. This approach has presented a limit of detection (LOD) of 25 RNA copies /reaction mix (1 copy/μL) and the test of cross-reactivity with other related viruses has not shown any cross-reactivity. Twenty clinical samples were evaluated by NALFA and simultaneously compared with the gold standard RT-qPCR protocol, originating equal results. Although the number of clinical specimens tested being relatively small, this indicates a sensitivity and specificity both of 100%. In short, an alternative NALFA was successfully implemented, rendering an accurate route for SARS-CoV-2 diagnosis, compatible with low-resource settings.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Vindeirinho et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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0 (RNA, Viral)
0 (DNA, Single-Stranded)
0 (DNA Primers)
0 (DNA Probes)

Date Created: 20240510 Date Completed: 20240510 Latest Revision: 20240512

20240512

PMC11086916

10.1371/journal.pone.0301234

38728290