Quantitative Ebola Virus RNA Detection

Viral RNA was quantified from serum as well as tissue samples via quantitative reverse transcription-PCR (qRT-PCR) using Applied Biosystems Quant Studio 3 real-time PCR instrument (Life Technologies Corporation, Carlsbad, CA, USA). Serum was inactivated using RNAbee (Tel-Test, Friendswood, TX, USA) and RNA was isolated according to the manufacturer’s instructions. One-step qRT-PCR was performed using RNA UltraSense One-Step Quantitative RT-PCR System (Invitrogen, Carlsbad, CA, USA), and primers and probe specifically designed to detect a region of the EBOV glycoprotein gene [26 (link)]; this One-Step method and primers will detect all RNA corresponding to the Ebola virus GP gene (including antigenome and mRNA).