The outbreak of coronavirus disease 2019 (COVID-19) has been declared a pandemic by the World Health Organization (WHO) and has resulted in the worst public health crisis since World War II (Wang et al. 2020). The causative pathogen of COVID-19 is a novel strain of coronavirus named severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2); SARS-CoV-2 is the seventh coronavirus that has been found to infect humans (Gorbalenyaet al. 2020; Zhang and Holmes 2020; Zhu et al. 2020). As of June 8, 2020, more than 7 million confirmed cases of COVID-19 have been reported worldwide, resulting in approximately 400,000 deaths (WHO 2020). As recommended by the WHO, detection and isolation are essential for containing the rapid spread of this pandemic. Therefore, adequate, large-scale screening and detection of SARS-CoV-2 are necessary. In addition, in locations in which the population has experienced and gradually recovered from COVID-19, there may be a huge demand for large-scale testing and surveillance. For example, in Wuhan city, in which the virus was successfully contained after its rapid spread, residents gradually resumed normal activities beginning in mid-March, with full re-opening of the economy by April 8, 2020. Data from Wuhan Municipal Health Commission showed that approximately 275,400 nucleic acid tests for SARS-CoV-2 had been conducted for people who returned to work in Wuhan from April 8 to April 15 (CCTV 2020). Therefore, all countries and cities, including those still experiencing outbreaks and those in the process of recovery, are expected to require mass COVID-19 testing, which could be limited by worldwide testing capacity. Currently, methods for detection of SARS-CoV-2 include SARS-CoV-2 nucleic acid detection, antibody (IgM/IgG) detection, and antigen detection (Corman et al. 2020; Seo et al. 2020; Xiang et al. 2020). Among these methods, nucleic acid detection is the most commonly used; the presence of viral RNAs is an indication of ongoing viral infection in the human body. At present, viral nucleic acids are generally sampled using throat swabs and detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). However, the capacity for SARS-CoV-2 nucleic acid detection is largely limited by the number of instruments, kits, and experienced laboratory personnel available. These limitations have caused low screening efficiency, which leads to a lag in identifying potential infections and may then exacerbate the spread of COVID-19 (Carter et al. 2020). Therefore, novel qRT-PCR approaches for nucleic acid detection are required to enhance testing efficiency.

In this study, we aimed to explore a practical method and procedure for SARS-CoV-2 qRT-PCR detection in 96-well plates using pooled throat swab samples. This method may reduce the cost of testing and increase the screening capacity for SARS-CoV-2 using existing instrument and kits, consequently facilitating the containment of COVID-19 worldwide. The pooled sample strategy was designed owing to the low detection of infection. A similar sample pooling strategy is often used in blood donation, and a recent report on SARS-CoV-2 from the United States of America described a similar approach (Hoganet al. 2020). Because the virus from positive samples will be diluted by negative samples, the detection kit must be highly