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Chow, Franklin Wang-Ngai; Chan, Tony Tat-Yin; Tam, Anthony Raymond; Zhao, Suhui; Yao, Weiming; Fung, Joshua; Cheng, Flora Ka-Kei; Lo, George Chi-Shing; Chu, Stella; Aw-Yong, Kam Leng; Tang, James Yat-Man; Tsang, Chi-Ching; Luk, Hayes Kam-Hei; Wong, Antonio Cheuk-Pui; Li, Kenneth Sze-Ming; Zhu, Longchao; He, Zirong; Tam, Emily Wan Ting; Chung, Tom Wai-Hin; Wong, Sally Cheuk Ying; Que, Tak-Lun; Fung, Kitty Sau-Chun; Lung, David Christopher; Wu, Alan Ka-Lun; Hung, Ivan Fan-Ngai; Woo, Patrick Chiu-Yat; Lau, Susanna Kar-Pui
International journal of molecular sciences, 07/2020, Volume: 21, Issue: 15Journal Article
To control the COVID-19 pandemic and prevent its resurgence in areas preparing for a return of economic activities, a method for a rapid, simple, and inexpensive point-of-care diagnosis and mass screening is urgently needed. We developed and evaluated a one-step colorimetric reverse-transcriptional loop-mediated isothermal amplification assay (COVID-19-LAMP) for detection of SARS-CoV-2, using SARS-CoV-2 isolate and respiratory samples from patients with COVID-19 ( = 223) and other respiratory virus infections ( = 143). The assay involves simple equipment and techniques and low cost, without the need for expensive qPCR machines, and the result, indicated by color change, is easily interpreted by naked eyes. COVID-19-LAMP can detect SARS-CoV-2 RNA with detection limit of 42 copies/reaction. Of 223 respiratory samples positive for SARS-CoV-2 by qRT-PCR, 212 and 219 were positive by COVID-19-LAMP at 60 and 90 min (sensitivities of 95.07% and 98.21%) respectively, with the highest sensitivities among nasopharyngeal swabs (96.88% and 98.96%), compared to sputum/deep throat saliva samples (94.03% and 97.02%), and throat swab samples (93.33% and 98.33%). None of the 143 samples with other respiratory viruses were positive by COVID-19-LAMP, showing 100% specificity. Samples with higher viral load showed shorter detection time, some as early as 30 min. This inexpensive, highly sensitive and specific COVID-19-LAMP assay can be useful for rapid deployment as mobile diagnostic units to resource-limiting areas for point-of-care diagnosis, and for unlimited high-throughput mass screening at borders to reduce cross-regional transmission.
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