Author: Anne Louise Wyllie; John Fournier; Arnau Casanovas-Massana; Melissa Campbell; Maria Tokuyama; Pavithra Vijayakumar; Bertie Geng; M. Catherine Muenker; Adam J. Moore; Chantal B. F. Vogels; Mary E. Petrone; Isabel M. Ott; Peiwen Lu; Alice Lu-Culligan; Jonathan Klein; Arvind Venkataraman; Rebecca Earnest; Michael Simonov; Rupak Datta; Ryan Handoko; Nida Naushad; Lorenzo R. Sewanan; Jordan Valdez; Elizabeth B. White; Sarah Lapidus; Chaney C. Kalinich; Xiaodong Jiang; Daniel J. Kim; Eriko Kudo; Melissa Linehan; Tianyang Mao; Miyu Moriyama; Ji Eun Oh; Annsea Park; Julio Silva; Eric Song; Takehiro Takahashi; Manabu Taura; Orr-El Weizman; Patrick Wong; Yexin Yang; Santos Bermejo; Camila Odio; Saad B. Omer; Charles S. Dela Cruz; Shelli Farhadian; Richard A. Martinello; Akiko Iwasaki; Nathan D. Grubaugh; Albert I. Ko
Title: Saliva is more sensitive for SARS-CoV-2 detection in COVID-19 patients than nasopharyngeal swabs Document date: 2020_4_22
ID: lt7qsxxh_26
Snippet: On arrival at the research lab, total nucleic acid was extracted from 300 µl of viral transport media from the nasopharyngeal swab or 300 µl of whole saliva using the MagMAX Viral/Pathogen Nucleic Acid Isolation kit (ThermoFisher Scientific) following the manufacturer's protocol and eluted into 75 µl of elution buffer. For SARS-CoV-2 RNA detection, 5 µl of RNA template was tested as previously described 21, 22 , using the US CDC real-time RT-.....
Document: On arrival at the research lab, total nucleic acid was extracted from 300 µl of viral transport media from the nasopharyngeal swab or 300 µl of whole saliva using the MagMAX Viral/Pathogen Nucleic Acid Isolation kit (ThermoFisher Scientific) following the manufacturer's protocol and eluted into 75 µl of elution buffer. For SARS-CoV-2 RNA detection, 5 µl of RNA template was tested as previously described 21, 22 , using the US CDC real-time RT-PCR primer/probe sets for 2019-nCoV_N1 and 2019-nCoV_N2 and the human RNase P (RP) as an extraction control. Samples were classified as positive for SARS-CoV-2 when both N1 and N2 primer-probe sets were detected <38 C T . Virus copies were quantified using a 10-fold dilution standard curve of RNA transcripts that we previously generated 21 . As results from N1 and N2 were comparable ( Extended Data Fig. 1 ), all virus copies are shown as calculated using the N1 primer-probe set.
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