One-step RNA extraction for RT-qPCR detection of 2019-nCoV

Monica Sentmanat; Evguenia Kouranova; Xiaoxia Cui

In the CDC protocol, specimens are typically placed in 3 mL of viral transport media consisting of a buffered salt solution with fetal bovine serum and an antimicrobial solution 2 . Viral particles in the samples remain infectious until lysed during RNA extraction. The RNA is then prepared by using a column based RNA extraction kit. We showed previously that RNA can be extracted by one-step lysis in QuickExtract DNA Extraction Solution (QE buffer) and directly used in RT-qPCR 3 . The QE buffer contains detergents and proteinase K, both of which could inactivate viral particles. Previous work on hepatitis C and ebola virus has shown that detergent alone is sufficient to reduce infectious titer in the absence of serum (e.g. fetal bovine serum) and is even more effective in combination with proteinase K which degrades core viral proteins accessible through lipid viral envelop dissolution by detergent 4, 5 .