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Direct clinical evidence recommending the use of Proteinase K or Dithiothreitol to pretreat sputum for detection of SARS-CoV-2 Peng, Jing; Lu, Yanjun; Song, Juan; Vallance, Bruce A.; Jacobson, Kevan; Yu, Hong Bing; Sun, Ziyong
Abstract
One of the primary tools for diagnosing COVID-19 is the nucleic acid-based real-time reverse transcriptase-polymerase chain reaction (RT-PCR) test performed on respiratory specimens. The detection rate of SARS-CoV-2 in lower respiratory specimens (such as sputum) is higher than that for upper respiratory specimens (such as nasal and pharyngeal swabs). However, sputum specimens are usually quite viscous, requiring a homogenization process prior to nucleic acid (NA) extraction for RT-PCR. Sputum specimens from COVID-19 and non-COVID-19 patients were treated with four commonly used reagents—saline, N-acetyl-L-cysteine (NALC), proteinase K (PK), and dithiothreitol (DTT), prior to NA extraction. These reagents were then compared for their performance in diagnosing COVID-19 in real clinical practice. The detection rate of SARS-CoV-2 in PK- or DTT-treated sputum was comparable, and higher than that in sputum treated with NALC or saline. While there was a 4.8% (1/21) false negative rate for the PK- and DTT-treated sputum, neither treatment showed any false positive cases among patients with non-COVID diseases. Moreover, sputum pretreated with saline, NALC, PK or DTT showed higher detection rates of SARS-CoV-2 as compared to pharyngeal swabs. Taken together, we provide direct evidence recommending the use of PK or DTT to pretreat sputum samples to facilitate SARS-CoV-2 detection by clinical laboratories. Moreover, our methods should help to standardize the procedure of processing sputum specimens and improve the ability to detect SARS-CoV-2 in these samples.
Item Metadata
Title |
Direct clinical evidence recommending the use of Proteinase K or Dithiothreitol to pretreat sputum for detection of SARS-CoV-2
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Creator | |
Contributor | |
Publisher |
Frontiers
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Date Issued |
2020-09-18
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Description |
One of the primary tools for diagnosing COVID-19 is the nucleic acid-based real-time
reverse transcriptase-polymerase chain reaction (RT-PCR) test performed on respiratory
specimens. The detection rate of SARS-CoV-2 in lower respiratory specimens (such
as sputum) is higher than that for upper respiratory specimens (such as nasal and
pharyngeal swabs). However, sputum specimens are usually quite viscous, requiring
a homogenization process prior to nucleic acid (NA) extraction for RT-PCR. Sputum
specimens from COVID-19 and non-COVID-19 patients were treated with four commonly
used reagents—saline, N-acetyl-L-cysteine (NALC), proteinase K (PK), and dithiothreitol
(DTT), prior to NA extraction. These reagents were then compared for their performance
in diagnosing COVID-19 in real clinical practice. The detection rate of SARS-CoV-2 in
PK- or DTT-treated sputum was comparable, and higher than that in sputum treated
with NALC or saline. While there was a 4.8% (1/21) false negative rate for the PK- and
DTT-treated sputum, neither treatment showed any false positive cases among patients
with non-COVID diseases. Moreover, sputum pretreated with saline, NALC, PK or DTT
showed higher detection rates of SARS-CoV-2 as compared to pharyngeal swabs. Taken
together, we provide direct evidence recommending the use of PK or DTT to pretreat
sputum samples to facilitate SARS-CoV-2 detection by clinical laboratories. Moreover,
our methods should help to standardize the procedure of processing sputum specimens
and improve the ability to detect SARS-CoV-2 in these samples.
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Subject | |
Genre | |
Type | |
Language |
eng
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Date Available |
2021-04-16
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Provider |
Vancouver : University of British Columbia Library
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Rights |
Attribution 4.0 International
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DOI |
10.14288/1.0396748
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URI | |
Affiliation | |
Citation |
Peng, J., Lu, Y., Song, J., Vallance, B. A., Jacobson, K., Yu, H. B., & Sun, Z. (2020). Direct clinical evidence recommending the use of proteinase K or dithiothreitol to pretreat sputum for detection of SARS-CoV-2. Frontiers in Medicine, 7.
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Publisher DOI |
10.3389/fmed.2020.549860
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Peer Review Status |
Reviewed
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Scholarly Level |
Faculty; Researcher
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Copyright Holder |
Authors
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Rights URI | |
Aggregated Source Repository |
DSpace
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Rights
Attribution 4.0 International