Direct Clinical Evidence Recommending the Use of Proteinase K or Dithiothreitol to Pretreat Sputum for Detection of SARS-CoV-2
| Autor: | Ziyong Sun, Juan Song, Jing Peng, Yanjun Lu, Bruce A. Vallance, Hongbing Yu, Kevan Jacobson |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
medicine.medical_specialty Coronavirus disease 2019 (COVID-19) Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) medicine.medical_treatment Gastroenterology Dithiothreitol 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Internal medicine medicine 030212 general & internal medicine Respiratory system Saline lcsh:R5-920 biology business.industry sputum COVID-19 General Medicine Brief Research Report Proteinase K proteinase K (PK) respiratory tract diseases 030104 developmental biology chemistry Clinical evidence biology.protein Medicine Sputum detection of SARS-CoV-2 medicine.symptom business lcsh:Medicine (General) DTT |
| Zdroj: | Frontiers in Medicine, Vol 7 (2020) Frontiers in Medicine |
| DOI: | 10.3389/fmed.2020.549860/full |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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