Author: Arunkumar Arumugam; Season S Wong
Title: The Potential Use of Unprocessed Sample for RT-qPCR Detection of COVID-19 without an RNA Extraction Step Document date: 2020_4_8
ID: ka3n9pft_9
Snippet: As shown in Fig. 3 , the SARS-CoV-2 RNA from directly spiked samples was successfully detected by the RT-qPCR reaction without a nucleic acid extraction step (N1 target shown). Up to 8 µL of VTM and AccuPlex SARS-CoV-2 mix was used in the reaction, which did not exert major inhibition on PCR amplification. The average Ct values are 38.5 (for 2 µL), 37.6 (for 4 µL), 38 (for 6 µL), and 39.1 (for 8 µL), still less than the 40 cycles that is con.....
Document: As shown in Fig. 3 , the SARS-CoV-2 RNA from directly spiked samples was successfully detected by the RT-qPCR reaction without a nucleic acid extraction step (N1 target shown). Up to 8 µL of VTM and AccuPlex SARS-CoV-2 mix was used in the reaction, which did not exert major inhibition on PCR amplification. The average Ct values are 38.5 (for 2 µL), 37.6 (for 4 µL), 38 (for 6 µL), and 39.1 (for 8 µL), still less than the 40 cycles that is considered to be a common cut- template (20 copies/reaction) is 35.5, which is 2 cycles lower than the result from 4 µL of untreated sample (10 copies) in the reaction (Fig. 3) . We speculate that the SeraCare matrix, containing Trisbuffered saline, glycerol, anti-microbial agents and human proteins, could be the reason for the above noticed PCR inhibition.
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