+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus



Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus



Plos Currents 5:



The emergence of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) in the eastern Mediterranean and imported cases to Europe has alerted public health authorities. Currently, detection of MERS-CoV in patient samples is done by real-time RT-PCR. Samples collected from suspected cases are sent to highly-equipped centralized laboratories for screening. A rapid point-of-care test is needed to allow more widespread mobile detection of the virus directly from patient material. In this study, we describe the development of a reverse transcription isothermal Recombinase Polymerase Amplification (RT-RPA) assay for the identification of MERS-CoV. A partial nucleocapsid gene RNA molecular standard of MERS-coronavirus was used to determine the assay sensitivity. The isothermal (42°C) MERS-CoV RT-RPA was as sensitive as real-time RT-PCR (10 RNA molecules), rapid (3-7 minutes) and mobile (using tubescanner weighing 1kg). The MERS-CoV RT-RPA showed cross-detection neither of any of the RNAs of several coronaviruses and respiratory viruses affecting humans nor of the human genome. The developed isothermal real-time RT-RPA is ideal for rapid mobile molecular MERS-CoV monitoring in acute patients and may also facilitate the search for the animal reservoir of MERS-CoV.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 055556590

Download citation: RISBibTeXText

PMID: 24459611


Related references

Reverse transcription recombinase polymerase amplification assay for the rapid detection of type 2 porcine reproductive and respiratory syndrome virus. Journal of Virological Methods 243: 55-60, 2017

Detection of Middle East Respiratory Syndrome Coronavirus by Reverse-transcription Loop-Mediated Isothermal Amplification. Bing du Xue Bao 31(3): 269-275, 2015

Detection of Middle East respiratory syndrome coronavirus using reverse transcription loop-mediated isothermal amplification (RT-LAMP). Virology Journal 11: 139, 2014

Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus. Journal of Virological Methods 258: 41-48, 2018

A new approach for diagnosis of bovine coronavirus using a reverse transcription recombinase polymerase amplification assay. Journal of Virological Methods 193(2): 337-340, 2013

Development of a reverse transcription recombinase polymerase amplification assay for rapid detection of human respiratory syncytial virus. Molecular and Cellular Probes 45: 8, 2019

Evaluation of a Real-Time Reverse Transcription-PCR (RT-PCR) Assay for Detection of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) in Clinical Samples from an Outbreak in South Korea in 2015. Journal of Clinical Microbiology 55(8): 2554-2555, 2017

Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus. Journal of Clinical Microbiology 52(1): 67-75, 2014

A Field-Deployable Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of the Chikungunya Virus. Plos Neglected Tropical Diseases 10(9): E0004953, 2016

Development of reverse transcription recombinase polymerase amplification assay for avian influenza H5N1 HA gene detection. Journal of Virological Methods 223: 45-49, 2015

Clinical validation of 3 commercial real-time reverse transcriptase polymerase chain reaction assays for the detection of Middle East respiratory syndrome coronavirus from upper respiratory tract specimens. Diagnostic Microbiology and Infectious Disease 87(4): 320-324, 2017

Development of a reverse transcription recombinase polymerase amplification assay for rapid detection of Theiler's murine encephalomyelitis virus. Molecular and Cellular Probes 41: 27-31, 2018

A portable reverse transcription recombinase polymerase amplification assay for rapid detection of foot-and-mouth disease virus. Plos one 8(8): E71642, 2013

Use of a rapid reverse-transcription recombinase aided amplification assay for respiratory syncytial virus detection. Diagnostic Microbiology and Infectious Disease 90(2): 90-95, 2018

Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus. Journal of Molecular Diagnostics 19(6): 817-827, 2017