Author: Minghua Jiang; Wenjie Fang; Amir Aratehfar; Xiaojing Li; Liyan ling; Hua Fang; Farnaz Farnaz Daneshnia; Jian Yu; Wanqing Liao; Hao Pei; Weihua Pan; Cornelia Lass-Florl
Title: Development and validation of a rapid single-step reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) system potentially to be used for reliable and high-throughput screening of COVID-19 Document date: 2020_3_20
ID: eo2pcgix_9
Snippet: Analytical sensitivity and specificity testing was performed in a P2 lab and in order to mimic the real 90 viral particles we purchased pseudotyped SARS-CoV-2 assay system containing ORF1ab part 91 sequence, N gene and E gene (DAAN gene Co. Ltd, Guangzhou, China). RNA of pseudotyped virus 92 were extracted using EZ-10 Spin Column Viral Total RNA Extraction Kit (Sangon Biotech Co.,Ltd. 93 Shanghai, China). Serial dilutions with the magnitude of lo.....
Document: Analytical sensitivity and specificity testing was performed in a P2 lab and in order to mimic the real 90 viral particles we purchased pseudotyped SARS-CoV-2 assay system containing ORF1ab part 91 sequence, N gene and E gene (DAAN gene Co. Ltd, Guangzhou, China). RNA of pseudotyped virus 92 were extracted using EZ-10 Spin Column Viral Total RNA Extraction Kit (Sangon Biotech Co.,Ltd. 93 Shanghai, China). Serial dilutions with the magnitude of log10 containing 50*10 6 cell/ml to 50*10 0 94 cell/ml pseudotyped virus were performed to determine the limit of detection (LOD). Serial dilution 95 testing was performed in both RNase/DNase free molecular grade water and sputum sample collected 96 from a COVID-19 negative healthy individual. Reproducibility of our LAMP assay (linearity=R 2 97 value) was assessed by separate serial dilution testing on three occasions, each performed in duplicate. 98 All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity.
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