4 Examples of RT-PCR diagnostic kits based on the aforementioned genes that are currently used in South Korea and the United States are listed in Supplementary 1 (Supplemental Digital Content 1, ). These countries have published their molecular diagnostic protocols and the primer/probe sequences on the World Health Organization website. Examples of a few countries and the target genes assayed are as follows: China (ORF1 ab, N), Germany (RdRP, E, N), Hong Kong (OLRF1b-nsp14, N), Japan (Pancoronavirus and multiple targets, S), Thailand (N), the United States (three targets in N), and France (two targets in RdRP). Most countries currently use rRT-PCR-based assays for the detection of COVID-19 infection. 3 Among them, the genes for the N and E proteins are used as the targets for amplification in the rRT-PCR assay combined with the open reading frame 1 (ORF1) ab, and the RNA-dependent RNA polymerase (RdRP) gene. The spike surface glycoprotein (S) mediates specific binding to the host cell receptors, the nucleocapsid (N) protein binds to the coronavirus RNA genome to make the nucleocapsid, the membrane (M) protein is the main structural protein that connects between the membrane and the capsid, and the small envelope (E) protein which is involved in the assembly and budding process of the coronavirus. The SARS-CoV-2 genome encodes four structural proteins. To understand the principle of the assay and the choice of primer sets used, some basic knowledge of COVID-19 biology is necessary. Currently, rRT-PCR is the most widely used diagnostic method for COVID-19.
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