id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-349623-dw5o9i59	Miranda, José P.	Analytical and Clinical Validation for RT-qPCR Detection of SARS-CoV-2 Without RNA Extraction	2020-10-15		.txt	text/plain	3813	174	48	Methods: Optimal direct protocol was selected by comparing RT-qPCR performance under a set of thermal (65, 70, and 95° for 5, 10, and 30 min) and amplification conditions (3 or 3.5 uL loading volume; 2 commercial RT-qPCR kits with a limit of detection below 10 copies/reaction) in nasopharyngeal swabs stored at 4°C in sterile Weise's buffer pH 7.2. For the standard protocol, routinely used in the laboratory for the detection of SARS-CoV-2, an aliquot of 180 ul of the sample from the nasopharyngeal swab, including 10 ul of extraction control, was used to extract RNA with the MagNA Pure 96 DNA and Viral NA LV Kit (Roche Diagnostics, Cat. No. For the standardization of the direct SARS-CoV-2 detection protocol without RNA extraction steps, 50 ul aliquots from the primary sample (nasopharyngeal swabs) of 5 anonymized patients were subjected to heat shock (65, 70, or 95 • C) during different incubation times (5, 10, or 30 min), and then were quickly placed at 4 • C until the moment of amplification.	./cache/cord-349623-dw5o9i59.txt	./txt/cord-349623-dw5o9i59.txt