单位：[1]Department of Laboratory Medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,China,2 Department of Gastroenterology & Endocrinology,Wuhan No. 9 Hospital,Wuhan,China华中科技大学同济医学院附属同济医院检验科内科学系内分泌内科消化内科[2]Department of Gastroenterology & Endocrinology, Wuhan No. 9 Hospital, Wuhan, China,[3]Department of Pediatrics,BC Children’s Hospital Research Institute,University of British Columbia,Vancouver,BC,Canada儿科学系
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.
基金：
National Mega Project on Major Infectious Disease Prevention [2017ZX10103005-007-001, 2017ZX10103005-007-002]
第一作者单位：[1]Department of Laboratory Medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,China,2 Department of Gastroenterology & Endocrinology,Wuhan No. 9 Hospital,Wuhan,China
共同第一作者：
通讯作者：
推荐引用方式(GB/T 7714)：
peng jing,lu yanjun,song juan,et al.Direct Clinical Evidence Recommending the Use of Proteinase K or Dithiothreitol to Pretreat Sputum for Detection of SARS-CoV-2[J].FRONTIERS IN MEDICINE.2020,7:doi:10.3389/fmed.2020.549860.
APA：
peng,jing,lu,yanjun,song,juan,vallance,bruce a.,jacobson,kevan...&sun,ziyong.(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,
MLA：
peng,jing,et al."Direct Clinical Evidence Recommending the Use of Proteinase K or Dithiothreitol to Pretreat Sputum for Detection of SARS-CoV-2".FRONTIERS IN MEDICINE 7.(2020)
