Abstract
Loop-mediated isothermal amplification (LAMP) is being used as a robust rapid diagnostic tool to prevent the transmission of infectious diseases. However, carryover contamination of LAMP-amplified products originating from previous tests has been a problem in LAMP-based bio-analytical assays. In this study, we developed a Cod-uracil-DNA-glycosylase real-time reverse transcriptase LAMP assay (Cod-UNG-rRT-LAMP) for the elimination of carryover contamination and the rapid detection of SARS-CoV-2 in point-of-care (POC) testing. Using the Cod-UNG-rRT-LAMP assay, the SARS-CoV-2 virus could be detected as low as 2 copies/µl (8 copies/reaction) within 45 min of amplification and 2.63 ± 0.17 pg (equivalent to 2.296 × 10 9 copies) of contaminants per reaction could be eliminated. Analysis of clinical SARS-CoV-2 samples using the Cod-UNG-rRT-LAMP assay showed an excellent agreement with a relative accuracy of 98.2%, sensitivity of 97.1%, and specificity of 95.2% in comparison to rRT-PCR. The results obtained in this study clearly demonstrate the feasibility of the use of the Cod-UNG-rRT-LAMP assay for applications toward the POC diagnosis of SARS-CoV-2 and on-site testing of other pathogens.
Keywords: COVID-19; Cod uracil DNA glycosylase (Cod-UNG); SARS-CoV-2; carryover contamination; loop-mediated isothermal amplification (LAMP); on-site testing; point-of-care.
【저자키워드】 COVID-19, SARS-CoV-2, point-of-care, Loop-mediated isothermal amplification (LAMP), Cod uracil DNA glycosylase (Cod-UNG), carryover contamination, on-site testing, 【초록키워드】 Infectious diseases, Diseases, feasibility, POC, diagnostic, SARS-CoV-2 virus, Transmission, Elimination, point-of-care, amplification, DNA, sensitivity, specificity, rRT-PCR, Contamination, Accuracy, Loop-mediated isothermal amplification, LAMP, Pathogens, isothermal amplification, reverse transcriptase, reaction, problem, uracil, COD, Prevent, robust, contaminant, real-time reverse transcriptase, glycosylase, assays, diagnosis of SARS-CoV-2, eliminated, the SARS-CoV-2 virus, 【제목키워드】 detection, Testing, Rapid, the SARS-CoV-2,