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      Stability issues of RT-PCR testing of SARS-CoV-2 for hospitalized patients clinically diagnosed with COVID-19.

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          Abstract

          In this study, we collected a total of 610 hospitalized patients from Wuhan between February 2, 2020, and February 17, 2020. We reported a potentially high false negative rate of real-time reverse-transcriptase polymerase chain reaction (RT-PCR) testing for SARS-CoV-2 in the 610 hospitalized patients clinically diagnosed with COVID-19 during the 2019 outbreak. We also found that the RT-PCR results from several tests at different points were variable from the same patients during the course of diagnosis and treatment of these patients. Our results indicate that in addition to the emphasis on RT-PCR testing, clinical indicators such as computed tomography images should also be used not only for diagnosis and treatment but also for isolation, recovery/discharge, and transferring for hospitalized patients clinically diagnosed with COVID-19 during the current epidemic. These results suggested the urgent needs for the standard of procedures of sampling from different anatomic sites, sample transportation, optimization of RT-PCR, serology diagnosis/screening for SARS-CoV-2 infection, and distinct diagnosis from other respiratory diseases such as fluenza infections as well.

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          Is Open Access

          Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR

          Background The ongoing outbreak of the recently emerged novel coronavirus (2019-nCoV) poses a challenge for public health laboratories as virus isolates are unavailable while there is growing evidence that the outbreak is more widespread than initially thought, and international spread through travellers does already occur. Aim We aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available. Methods Here we present a validated diagnostic workflow for 2019-nCoV, its design relying on close genetic relatedness of 2019-nCoV with SARS coronavirus, making use of synthetic nucleic acid technology. Results The workflow reliably detects 2019-nCoV, and further discriminates 2019-nCoV from SARS-CoV. Through coordination between academic and public laboratories, we confirmed assay exclusivity based on 297 original clinical specimens containing a full spectrum of human respiratory viruses. Control material is made available through European Virus Archive – Global (EVAg), a European Union infrastructure project. Conclusion The present study demonstrates the enormous response capacity achieved through coordination of academic and public laboratories in national and European research networks.
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            Chest CT for Typical 2019-nCoV Pneumonia: Relationship to Negative RT-PCR Testing

            Some patients with positive chest CT findings may present with negative results of real time reverse-transcription–polymerase chain- reaction (RT-PCR) for 2019 novel coronavirus (2019-nCoV). In this report, we present chest CT findings from five patients with 2019-nCoV infection who had initial negative RT-PCR results. All five patients had typical imaging findings, including ground-glass opacity (GGO) (5 patients) and/or mixed GGO and mixed consolidation (2 patients). After isolation for presumed 2019-nCoV pneumonia, all patients were eventually confirmed with 2019-nCoV infection by repeated swab tests. A combination of repeated swab tests and CT scanning may be helpful when for individuals with high clinical suspicion of nCoV infection but negative RT-PCR screening
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              Use of Chest CT in Combination with Negative RT-PCR Assay for the 2019 Novel Coronavirus but High Clinical Suspicion

              Online supplemental material is available for this article. A 36-year-old man presented to the hospital with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China. His temperature on admission was 37.8°C (100.04°F). Pulmonary auscultation was normal. Laboratory studies showed a normal white blood cell count (4.6 × 109/L) with a differential count of 53.1% neutrophils. The blood procalcitonin level was normal. Chest CT showed multiple peripheral ground-glass opacities in both lungs with more involvement of the left upper lobe, lingular segment (Figure a–c). At admission, the real-time fluorescence polymerase chain reaction (RT-PCR) assay of the sputum was negative for the 2019 novel coronavirus (2019-nCoV) nucleic acid. Figure a: Images in a 36-year-old man with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China, and a negative sputum real-time fluorescence polymerase chain reaction assay for the 2019 novel coronavirus. (a, b) Chest CT scans obtained at presentation show ground-glass opacities (red box) in the right upper lobe and the lingular segment and left lower lobe (b). (c) Volume rendering of chest CT scan obtained at admission. (d, e) CT scans obtained 3 days after admission show progression of ground-glass opacities to an atoll sign in the right upper lobe (red boxes in d) and left lower lobe consolidation (red boxes in e). (f) Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation. See also Movies 1 and 2 (online) Figure b: Images in a 36-year-old man with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China, and a negative sputum real-time fluorescence polymerase chain reaction assay for the 2019 novel coronavirus. (a, b) Chest CT scans obtained at presentation show ground-glass opacities (red box) in the right upper lobe and the lingular segment and left lower lobe (b). (c) Volume rendering of chest CT scan obtained at admission. (d, e) CT scans obtained 3 days after admission show progression of ground-glass opacities to an atoll sign in the right upper lobe (red boxes in d) and left lower lobe consolidation (red boxes in e). (f) Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation. See also Movies 1 and 2 (online) Figure c: Images in a 36-year-old man with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China, and a negative sputum real-time fluorescence polymerase chain reaction assay for the 2019 novel coronavirus. (a, b) Chest CT scans obtained at presentation show ground-glass opacities (red box) in the right upper lobe and the lingular segment and left lower lobe (b). (c) Volume rendering of chest CT scan obtained at admission. (d, e) CT scans obtained 3 days after admission show progression of ground-glass opacities to an atoll sign in the right upper lobe (red boxes in d) and left lower lobe consolidation (red boxes in e). (f) Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation. See also Movies 1 and 2 (online) Figure d: Images in a 36-year-old man with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China, and a negative sputum real-time fluorescence polymerase chain reaction assay for the 2019 novel coronavirus. (a, b) Chest CT scans obtained at presentation show ground-glass opacities (red box) in the right upper lobe and the lingular segment and left lower lobe (b). (c) Volume rendering of chest CT scan obtained at admission. (d, e) CT scans obtained 3 days after admission show progression of ground-glass opacities to an atoll sign in the right upper lobe (red boxes in d) and left lower lobe consolidation (red boxes in e). (f) Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation. See also Movies 1 and 2 (online) Figure e: Images in a 36-year-old man with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China, and a negative sputum real-time fluorescence polymerase chain reaction assay for the 2019 novel coronavirus. (a, b) Chest CT scans obtained at presentation show ground-glass opacities (red box) in the right upper lobe and the lingular segment and left lower lobe (b). (c) Volume rendering of chest CT scan obtained at admission. (d, e) CT scans obtained 3 days after admission show progression of ground-glass opacities to an atoll sign in the right upper lobe (red boxes in d) and left lower lobe consolidation (red boxes in e). (f) Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation. See also Movies 1 and 2 (online) Figure f: Images in a 36-year-old man with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China, and a negative sputum real-time fluorescence polymerase chain reaction assay for the 2019 novel coronavirus. (a, b) Chest CT scans obtained at presentation show ground-glass opacities (red box) in the right upper lobe and the lingular segment and left lower lobe (b). (c) Volume rendering of chest CT scan obtained at admission. (d, e) CT scans obtained 3 days after admission show progression of ground-glass opacities to an atoll sign in the right upper lobe (red boxes in d) and left lower lobe consolidation (red boxes in e). (f) Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation. See also Movies 1 and 2 (online) Movie 1: Volume rendering of chest CT scan obtained at admission (see Figure c). Movie 2: Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation (see Figure f). Repeat CT chest performed 3 days after admission showed transformation of ground-glass opacities to more consolidation (Figure d–f). A repeat RT-PCR 2019-nCoV nucleic acid assay was also negative at this time. Six days after admission, the third RT-PCR 2019-nCoV nucleic acid assay was finally found to be positive. When specimen tests are negative, the possibility of a false-negative result should be considered in the context of a patient’s recent exposures and the presence of clinical signs and symptoms consistent with 2019-nCoV infection (1,2). In this case, chest CT findings were typical of findings for 2019-nCoV pneumonia (3) coupled with recent exposure suggesting that 2019-nCoV infection was likely.
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                Author and article information

                Journal
                J Med Virol
                Journal of medical virology
                Wiley
                1096-9071
                0146-6615
                July 2020
                : 92
                : 7
                Affiliations
                [1 ] Department of Intensive Care Unit, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
                [2 ] Department of Operation and Management, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
                [3 ] Natural Language Processing Group, iFLYTEK Research South China, Guangzhou, China.
                [4 ] Department of Clinical Medicine, Zhongshan School of Medicine, Guangzhou, China.
                [5 ] Department of Respiratory, Hankou Hospital of Wuhan, Wuhan, China.
                Article
                10.1002/jmv.25786
                7228231
                32219885
                c0efe3fe-277e-45ca-ae85-5e25d80215d8
                © 2020 Wiley Periodicals, Inc.
                History

                COVID-19,RT-PCR
                COVID-19, RT-PCR

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