About: 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

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About: 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 Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	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
Creator	Dorostkar, Ruhollah Izadi, Morteza Najarasl, Mohammad Rasouli, Rahimeh Tat, Mahdi Zahraei, Bentolhoda Hashemzadeh, Mohammad Nejad, Behzad Saadat, Seyed
Source	Medline; PMC
abstract	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 and final confirmation of MERS-CoV infection, as recommended by the world health 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 two assay signatures and controls was assembled, which can be distributed to public health laboratories in Iran to support international MERS-CoV surveillance and public health response.
has issue date	2016-06-21(xsd:dateTime)
bibo:doi	10.5812/ircmj.23874
bibo:pmid	28191331
has license	cc-by-nc
sha1sum (hex)	9f9984a45e51e8268ed00e6de9ca8b23615fb154
schema:url	https://doi.org/10.5812/ircmj.23874
resource representing a document's title	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
has PubMed Central identifier	PMC5292137
has PubMed identifier	28191331
schema:publication	Iran Red Crescent Med J
resource representing a document's body	covid:9f9984a45e51e8268ed00e6de9ca8b23615fb154#body_text
is schema:about of	named entity 'PCR ASSAY' named entity 'KIT' named entity 'CORONAVIRUSES' named entity 'RESPIRATORY DISEASE' named entity 'LARGE' named entity 'RELIABLE' named entity 'CAUSING' named entity 'acid' named entity 'MERS-CoV' named entity 'RNA' named entity 'Saudi Arabia' named entity 'Coronavirus' named entity 'TaqMan' named entity 'Assay' named entity 'STEP' named entity 'NOVEL' named entity 'DEATHS' named entity 'SAUDI ARABIA' named entity 'ACCURATE' named entity 'BASED' named entity 'TAQMAN' named entity 'HUMANS' named entity 'NOVEL' named entity 'DIAGNOSTIC TEST' named entity 'DUAL' named entity 'DEVELOPMENT' named entity 'MERS-COV' named entity 'RRT-PCR' named entity 'PANEL' named entity 'SEPTEMBER' named entity 'HUMAN CORONAVIRUS' named entity 'DEVELOPMENT' named entity 'SPREAD' named entity 'RAPID' named entity 'RAPID' named entity 'HUMAN CORONAVIRUS' named entity 'REPORTED' named entity 'NUMBERS' named entity 'BACKGROUND' named entity 'MIDDLE EAST RESPIRATORY SYNDROME CORONAVIRUS' named entity 'INFECTIONS' named entity 'INCREASING' named entity 'NAMED' named entity 'VIRUSES' named entity 'TO PREVENT' named entity 'RIBONUCLEIC ACID' covid:arg/9f9984a45e51e8268ed00e6de9ca8b23615fb154 named entity 'causing' named entity 'infections' named entity 'respiratory disease' named entity 'rapid' named entity 'RNA' named entity 'ribonucleic acid' named entity 'Diagnostic Test' named entity 'rRT-PCR' named entity 'Red Crescent' named entity 'MERS-CoV' named entity 'viral genome' named entity 'MERS-CoV' named entity 'primers' named entity 'gene' named entity 'severe infections' named entity 'world health organization' named entity 'TaqMan' named entity 'early-stage' named entity 'NCBI' named entity 'ribonucleic acid' named entity 'virus' named entity 'MERS-CoV' named entity 'real-time RT-PCR' named entity 'rRT-PCR' named entity 'Qiagen' named entity 'vector'

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