Double-Quencher Probes Improved the Detection Sensitivity of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) by One-Step RT-PCR

Yosuke Hirotsu; Hitoshi Mochizuki; Masao Omata

Human transmission of SARS-CoV-2 has become a global health concern. The molecular diagnosis of SARS-CoV-2 is highly prioritized to establish the prevention of the viral outbreak and treatment of infected patients. Therefore, demands in detecting the virus in suspected patients in hospitals and clinical laboratories are growing. The Japanese government announced the approval of real-time PCR analysis to detect SARS-CoV-2 for health insurance on March 6, 2020. Here, we compared the detection sensitivity among the primer/probe sets designed by the National Institute of Infectious Diseases (NIID) in Japan and the Centers for Disease Control and Prevention (CDC) in the United States of America. These probes were designed as single-quencher probes. We originally designed the double-quencher probes consisting of the same nucleotide sequencing imposed by the NIID. The results showed that double-quenching probes reduced the background signal and improved the detection sensitivity of SARS-CoV-2.