In:
PLOS Biology, Public Library of Science (PLoS), Vol. 18, No. 12 ( 2020-12-15), p. e3001030-
Abstract:
With the ongoing COVID-19 (Coronavirus Disease 2019) pandemic, caused by the novel coronavirus SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2), there is a need for sensitive, specific, and affordable diagnostic tests to identify infected individuals, not all of whom are symptomatic. The most sensitive test involves the detection of viral RNA using RT-qPCR (quantitative reverse transcription PCR), with many commercial kits now available for this purpose. However, these are expensive, and supply of such kits in sufficient numbers cannot always be guaranteed. We therefore developed a multiplex assay using well-established SARS-CoV-2 targets alongside a human cellular control ( RPP30 ) and a viral spike-in control (Phocine Herpes Virus 1 [PhHV-1]), which monitor sample quality and nucleic acid extraction efficiency, respectively. Here, we establish that this test performs as well as widely used commercial assays, but at substantially reduced cost. Furthermore, we demonstrat e 〉 1,000-fold variability in material routinely collected by combined nose and throat swabbing and establish a statistically significant correlation between the detected level of human and SARS-CoV-2 nucleic acids. The inclusion of the human control probe in our assay therefore provides a quantitative measure of sample quality that could help reduce false-negative rates. We demonstrate the feasibility of establishing a robust RT-qPCR assay at approximately 10% of the cost of equivalent commercial assays, which could benefit low-resource environments and make high-volume testing affordable.
Type of Medium:
Online Resource
ISSN:
1545-7885
DOI:
10.1371/journal.pbio.3001030
DOI:
10.1371/journal.pbio.3001030.g001
DOI:
10.1371/journal.pbio.3001030.g002
DOI:
10.1371/journal.pbio.3001030.g003
DOI:
10.1371/journal.pbio.3001030.g004
DOI:
10.1371/journal.pbio.3001030.t001
DOI:
10.1371/journal.pbio.3001030.t002
DOI:
10.1371/journal.pbio.3001030.t003
DOI:
10.1371/journal.pbio.3001030.s001
DOI:
10.1371/journal.pbio.3001030.s002
DOI:
10.1371/journal.pbio.3001030.s003
DOI:
10.1371/journal.pbio.3001030.s004
DOI:
10.1371/journal.pbio.3001030.s005
DOI:
10.1371/journal.pbio.3001030.s006
DOI:
10.1371/journal.pbio.3001030.s007
DOI:
10.1371/journal.pbio.3001030.s008
DOI:
10.1371/journal.pbio.3001030.s009
DOI:
10.1371/journal.pbio.3001030.s010
DOI:
10.1371/journal.pbio.3001030.s011
DOI:
10.1371/journal.pbio.3001030.s012
DOI:
10.1371/journal.pbio.3001030.s013
DOI:
10.1371/journal.pbio.3001030.s014
DOI:
10.1371/journal.pbio.3001030.s015
DOI:
10.1371/journal.pbio.3001030.s016
DOI:
10.1371/journal.pbio.3001030.s017
DOI:
10.1371/journal.pbio.3001030.r001
DOI:
10.1371/journal.pbio.3001030.r002
DOI:
10.1371/journal.pbio.3001030.r003
DOI:
10.1371/journal.pbio.3001030.r004
DOI:
10.1371/journal.pbio.3001030.r005
DOI:
10.1371/journal.pbio.3001030.r006
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2020
detail.hit.zdb_id:
2126773-X
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