Establishment of a new method for rapid detection of SARS-CoV-2 by visual LAMP (without nucleic acid extraction)

ABSTRACT

A new type of coronavirus (SARS-CoV-2) infection caused acute or fatal pneumonia. The virus is another coronavirus that is transmitted from animal to human and capable of transmitted from human to human, following the severe acute respiratory syndrome coronavirus (SARS-CoV) and the Middle East respiratory syndrome coronavirus (MERS CoV). In order to control the epidemic as soon as possible, there is an urgent need, for rapid detection and confirmation of infected patients. In this study, according to the SARS CoV-2 whole genome published in GenBank as target gene, LAMP Desiner 2.0 software was used to screen efficient and highly specific combinatorial loop primers. The amplification characteristics of Bst 4.0 DNA polymerase relys RNA as template for DNA synthesis. Viral RNA-positive test results showed that 5 to 20 copies of virus nucleic acid could be detected. The inactivated virus was directly used as amplification template for clinical detection. The amplified nucleic acid molecules are combined with OG (Orange-Green) dye. Positive samples are green and negative samples are orange yellow.. The established SARS-CoV-2 one-step visual constant temperature rapid detection method realizes rapid detection of nucleic acids with high sensitivity. This study provides a new method for SARS-CoV-2 detection.