فهرست نشریات علمی دسترسی آزاد ایران

Coronaviruses (CoVs) are large ribonucleic acid (RNA) viruses causing primarily respiratory disease in humans. Anovel human coronavirus, subsequently named middle east respiratory syndrome coronavirus (MERS-CoV), was first reported inSaudi Arabia in September of 2012. With increasing numbers of infections and deaths from MERS-CoV, development of a rapid andreliable kit was crucial to prevent further spread of MERS-CoV.Objectives: In this study, we present two real-time reverse-transcription polymerase chain reaction (rRT-PCR) assays for in-houserapid and sensitive diagnostic testing of MERS-CoV, detecting the regions upstream of the envelope gene (upE) and open readingframe (ORF) 1b, respectively, for initial screening andfinal confirmation of MERS-CoV infection, asrecommendedby the worldhealthorganization (WHO).Materials and Methods: In this experimental study, acquiring patient samples was difficult; thus, according to WHO recommendationsand standard protocols, we synthesized RNA sequences of upE and ORF1b genes as the template signatures and TaqManbased-diagnostic rRT-PCR assays were carried out using these synthetic genes for detection of MERS-CoV. In this research, we alsoinaugurated a cell-free system to transcribe these RNA sequences using the DNA templates synthesized.Results: The upE and ORF1b based one-step rRT-PCR assays were optimized by testing several times via different synthetic RNAs, andvalidation results were highly successful. The sensitivity obtained for upE was fewer than ten copies of RNA template per reactionand for ORF1b was 50 or fewer copies per reaction.Conclusions: This study showed that the developed rRT-PCR assays are rapid, reliable, reproducible, specific, sensitive, and simpletools for detection of MERS-CoV. Finally, a kit consisting of twoassay signaturesandcontrolswasassembled, whichcanbe distributedto public health laboratories in Iran to support international MERS-CoV surveillance and public health response.