Development of dual TaqMan based one-step rRT-PCR assay panel for rapid and accurate diagnostic test of MERS-CoV : a novel human coronavirus, ahead of hajj pilgrimage
Joint Authors
Hashemzadeh, Muhammad Sadegh
Rasouli, Rahimah
Zahraei, Bentolhoda
Tat, Mahdi
Saadat, Sayyid Hassan
Nejad, Behzad Khansari
Dorostkar, Ruhollah
Izadi, Morteza
Source
Iranian Red Crescent Medical Journal
Issue
Vol. 18, Issue 11 (30 Nov. 2016), pp.1-7, 7 p.
Publisher
Publication Date
2016-11-30
Country of Publication
United Arab Emirates
No. of Pages
7
Main Subjects
Pharmacy, Health & Medical Sciences
Abstract EN
Background : Coronaviruses (CoVs) are large ribonucleic acid (RNA) viruses causing primarily respiratory disease in humans.
A novel human coronavirus, subsequently named middle east respiratory syndrome coronavirus (MERS-CoV), was first reported in Saudi Arabia in September of 2012.
With increasing numbers of infections and deaths from MERS-CoV, development of a rapid and reliable 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-house rapid and sensitive diagnostic testing of MERS-CoV, detecting the regions upstream of the envelope gene (upE) and open reading frame (ORF) 1b, respectively, for initial screening andfinal confirmation of MERS-CoV infection, asrecommendedby the worldhealth organization (WHO).
Materials and Methods: In this experimental study, acquiring patient samples was difficult; thus, according to WHO recommendations and standard protocols, we synthesized RNA sequences of upE and ORF1b genes as the template signatures and TaqMan based-diagnostic rRT-PCR assays were carried out using these synthetic genes for detection of MERS-CoV.
In this research, we also inaugurated 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, and validation results were highly successful.
The sensitivity obtained for upE was fewer than ten copies of RNA template per reaction and 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 simple tools for detection of MERS-CoV.
Finally, a kit consisting of twoassay signaturesandcontrolswasassembled, whichcanbe distributed to public health laboratories in Iran to support international MERS-CoV surveillance and public health response.
American Psychological Association (APA)
Hashemzadeh, Muhammad Sadegh& Rasouli, Rahimah& Zahraei, Bentolhoda& Izadi, Morteza& Tat, Mahdi& Saadat, Sayyid Hassan…[et al.]. 2016. Development of dual TaqMan based one-step rRT-PCR assay panel for rapid and accurate diagnostic test of MERS-CoV : a novel human coronavirus, ahead of hajj pilgrimage. Iranian Red Crescent Medical Journal،Vol. 18, no. 11, pp.1-7.
https://search.emarefa.net/detail/BIM-728725
Modern Language Association (MLA)
Hashemzadeh, Muhammad Sadegh…[et al.]. Development of dual TaqMan based one-step rRT-PCR assay panel for rapid and accurate diagnostic test of MERS-CoV : a novel human coronavirus, ahead of hajj pilgrimage. Iranian Red Crescent Medical Journal Vol. 18, no. 11 (Nov. 2016), pp.1-7.
https://search.emarefa.net/detail/BIM-728725
American Medical Association (AMA)
Hashemzadeh, Muhammad Sadegh& Rasouli, Rahimah& Zahraei, Bentolhoda& Izadi, Morteza& Tat, Mahdi& Saadat, Sayyid Hassan…[et al.]. Development of dual TaqMan based one-step rRT-PCR assay panel for rapid and accurate diagnostic test of MERS-CoV : a novel human coronavirus, ahead of hajj pilgrimage. Iranian Red Crescent Medical Journal. 2016. Vol. 18, no. 11, pp.1-7.
https://search.emarefa.net/detail/BIM-728725
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references : p. 6-7
Record ID
BIM-728725