Effects of Different Temperature and Time Durations of Virus Inactivation on Results of Real-time Fluorescence PCR Testi
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40(4):1-4,2020
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Effects of Different Temperature and Time Durations of Virus Inactivation on Results of Real-time Fluorescence PCR Testing of COVID-19 Viruses* Ze-gang WU†, Hong-yun ZHENG†, Jian GU†, Feng LI, Rui-long LV, Ya-yun DENG, Wan-zhou XU, Yong-qing TONG# Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan 430060, China Huazhong University of Science and Technology 2020
Summary: The novel coronavirus SARS-CoV-2 caused an outbreak of pneumonia in Wuhan, Hubei province of China in January 2020. This study aims to investigate the effects of different temperature and time durations of virus inactivation on the results of PCR testing for SARSCoV-2. Twelve patients at the Renmin Hospital of Wuhan University suspected of being infected with SARS-CoV-2 were selected on February 13, 2020 and throat swabs were taken. The swabs were stored at room temperature (20–25°C), then divided into aliquots and subjected to different temperature for different periods in order to inactivate the viruses (56°C for 30, 45, 60 min; 65, 70, 80°C for 10, 15, 20 min). Control aliquots were stored at room temperature for 60 min. Then all aliquots were tested in a real-time fluorescence PCR using primers against SARS-CoV-2. Regardless of inactivation temperature and time, 7 of 12 cases (58.3%) tested were positive for SARS-CoV-2 by PCR, and cycle threshold values were similar. These results suggest that virus inactivation parameters exert minimal influence on PCR test results. Inactivation at 65°C for 10 min may be sufficient to ensure safe, reliable testing. Key words: SARS-CoV-2; COVID-19; throat swabs; real-time fluorescence PCR
Numerous infections of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) have occurred in Wuhan, Hubei since December 2019, and the number of infections has rapidly increased[1]. New infections have since occurred in other regions of China and the other countries[2–4]. The SARS-CoV-2 has been added to the Chinese National Health Committee’s list of Class A infectious diseases requiring prevention and control under the Infectious Disease Control Law. As of April 10, 2020, a total of 81 953 confirmed cases and 3339 deaths (4.07%) have been reported nationwide[5]. Since the beginning of the epidemic in late December 2019, SARS-CoV-2 has now spread to all continents, and up to April 10, 2020, the WHO reported 1 521 252 confirmed cases and 92 798 deaths globally (Situation Ze-gang WU, E-mail: [email protected]; Hongyun ZHENG, E-mail: [email protected]; Jian GU, E-mail: [email protected] † These authors contributed equally to this work. # Corresponding author, E-mail: [email protected] * This work was supported by grants from the Special Science and Technology Cooperation Project of Ningxia Hui Autonomous Region Key R&D Program (No. 2018BFG02008) and the National Science and Technology Key Projects on “Major Infectious Diseases such as HIV/ AIDS, Viral Hepatitis Prevention and Treatment” (No. 2017ZX10103005).
Report-81). Real-time fluorescence PCR detection of SARSCoV-2
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