In:
PLOS ONE, Public Library of Science (PLoS), Vol. 15, No. 12 ( 2020-12-31), p. e0244882-
Abstract:
SARS-CoV-2 testing is crucial to controlling the spread of this virus, yet shortages of nucleic acid extraction supplies and other key reagents have hindered the response to COVID-19 in the US. Several groups have described loop-mediated isothermal amplification (LAMP) assays for SARS-CoV-2, including testing directly from nasopharyngeal swabs and eliminating the need for reagents in short supply. Frequent surveillance of individuals attending work or school is currently unavailable to most people but will likely be necessary to reduce the ~50% of transmission that occurs when individuals are nonsymptomatic. Here we describe a fluorescence-based RT-LAMP test using direct nasopharyngeal swab samples and show consistent detection in clinically confirmed primary samples with a limit of detection (LOD) of ~625 copies/μl, approximately 100-fold lower sensitivity than qRT-PCR. While less sensitive than extraction-based molecular methods, RT-LAMP without RNA extraction is fast and inexpensive. Here we also demonstrate that adding a lysis buffer directly into the RT-LAMP reaction improves the sensitivity of some samples by approximately 10-fold. Furthermore, purified RNA in this assay achieves a similar LOD to qRT-PCR. These results indicate that high-throughput RT-LAMP testing could augment qRT-PCR in SARS-CoV-2 surveillance programs, especially while the availability of qRT-PCR testing and RNA extraction reagents is constrained.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0244882
DOI:
10.1371/journal.pone.0244882.g001
DOI:
10.1371/journal.pone.0244882.g002
DOI:
10.1371/journal.pone.0244882.g003
DOI:
10.1371/journal.pone.0244882.g004
DOI:
10.1371/journal.pone.0244882.s001
DOI:
10.1371/journal.pone.0244882.s002
DOI:
10.1371/journal.pone.0244882.s003
DOI:
10.1371/journal.pone.0244882.s004
DOI:
10.1371/journal.pone.0244882.s005
DOI:
10.1371/journal.pone.0244882.r001
DOI:
10.1371/journal.pone.0244882.r002
DOI:
10.1371/journal.pone.0244882.r003
DOI:
10.1371/journal.pone.0244882.r004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2020
detail.hit.zdb_id:
2267670-3
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