In Vitro Synthesis of SFTSV RNAs

SFTSV RNAs were synthesized by in vitro transcription (IVT) reaction and used to determine the sensitivity and specificity of SFTSV DETECTR. The S gene fragments (nucleotides 1216 to 1370) of SFTSV genotypes A, B, C, D, E and F (GenBank accession numbers: KF791948, KP663745, AB985541, KP 663733, HQ141606 and KF358693, respectively) [5 (link)] were synthesized (Macrogen, Seoul, Korea) and amplified by PCR using primers containing a T7 promoter (S1 Table). IVT reaction was carried out with the PCR products by using the mMESSAGE mMACHINE T7 Transcription kit (Invitrogen, Waltham, MA, USA) according to the manufacturer’s instructions. The synthesized SFTSV RNAs were purified using RNA Clean and Concentrator 5 columns (Zymo research, Irvine, CA, USA).