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Basic Characteristics of Mutations
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Mutation Site
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A1493G |
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Mutation Site Sentence
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In silico analysis suggested that the N501Y-RT-LAMP and Q493R-RT-LAMP assays would distinguish N501Y (A1501U) and Q493R (A1478G)/Q498R (A1493G) strains from all coronaviruses lacking the target mutations (Supplementary Figure S3). |
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Mutation Level
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Nucleotide level |
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Mutation Type
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Nonsynonymous substitution |
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Gene/Protein/Region
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S |
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Standardized Encoding Gene
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S
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Genotype/Subtype
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BA.1;BA.2 |
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Viral Reference
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NC_045512.2
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Functional Impact and Mechanisms
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Disease
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COVID-19
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Immune
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- |
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Target Gene
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-
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Clinical and Epidemiological Correlations
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Clinical Information
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- |
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Treatment
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- |
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Location
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- |
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Literature Information
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PMID
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37445876
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Title
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A New Method to Detect Variants of SARS-CoV-2 Using Reverse Transcription Loop-Mediated Isothermal Amplification Combined with a Bioluminescent Assay in Real Time (RT-LAMP-BART)
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Author
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Iijima T,Sakai J,Kanamori D,Ando S,Nomura T,Tisi L,Kilgore PE,Percy N,Kohase H,Hayakawa S,Maesaki S,Hoshino T,Seki M
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Journal
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International journal of molecular sciences
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Journal Info
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2023 Jun 27;24(13):10698
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Abstract
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Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), of which there are several variants. The three major variants (Alpha, Delta, and Omicron) carry the N501Y, L452R, and Q493R/Q498R mutations, respectively, in the S gene. Control of COVID-19 requires rapid and reliable detection of not only SARS-CoV-2 but also its variants. We previously developed a reverse transcription loop-mediated isothermal amplification assay combined with a bioluminescent assay in real time (RT-LAMP-BART) to detect the L452R mutation in the SARS-CoV-2 spike protein. In this study, we established LAMP primers and peptide nucleic acid probes to detect N501Y and Q493R/Q498R. The LAMP primer sets and PNA probes were designed for the N501Y and Q493R/Q498R mutations on the S gene of SARS-CoV-2. The specificities of RT-LAMP-BART assays were evaluated using five viral and four bacterial reference strains. The sensitivities of RT-LAMP-BART assays were evaluated using synthetic RNAs that included the target sequences, together with RNA-spiked clinical nasopharyngeal and salivary specimens. The results were compared with those of conventional real-time reverse transcription-polymerase chain reaction (RT-PCR) methods. The method correctly identified N501Y and Q493R/Q498R. Within 30 min, the RT-LAMP-BART assays detected up to 100-200 copies of the target genes; conventional real-time RT-PCR required 130 min and detected up to 500-3000 copies. Surprisingly, the real-time RT-PCR for N501Y did not detect the BA.1 and BA.2 variants (Omicron) that exhibited the N501Y mutation. The novel RT-LAMP-BART assay is highly specific and more sensitive than conventional real-time RT-PCR. The new assay is simple, inexpensive, and rapid; thus, it can be useful in efforts to identify SARS-CoV-2 variants of concern.
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Sequence Data
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"EPI_ISL_601443 (Alpha), EPI_ISL_660190 (Beta), EPI_ISL_906081 (Gamma), EPI_ISL_1419152 (Delta), EPI_ISL_8750789 (Epsilon), EPI_ISL_8005633 (Zeta), EPI_ISL_1083809 (Eta), EPI_ISL_8158591 (Iota), EPI_ISL_8259884 (Mu), EPI_ISL_6841980 (Omicron BA.1), EPI_ISL_9097416 (Omicron BA.2), EPI_ISL_13241867 (Omicron BA.4), EPI_ISL_12703378 (Omicron BA.5)], SARS-CoV-1 (NC_004718.3), MERS-CoV (NC_019843.3)], and four human coronaviruses [α-coronaviruses AF304460.1 (229E) and AY567487.2 (NL63), and β-coronaviruses AY391777.1 (OC43) and NC_006577.2 (HKU1)"
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