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PLoS Currents Outbreaks. Reverse Transcription Recombinase Polymerase Amplification Assay for the Detection of Middle East Respiratory Syndrome Coronavirus
[Source: PLoS Currents Outbreaks, full page: (LINK). Abstract, edited.]
Reverse Transcription Recombinase Polymerase Amplification Assay for the Detection of Middle East Respiratory Syndrome Coronavirus
December 12, 2013 ? Research
Abd El Wahed A, Patel P, Heidenreich D, Hufert FT, Weidmann M. Reverse Transcription Recombinase Polymerase Amplification Assay for the Detection of Middle East Respiratory Syndrome Coronavirus. PLOS Currents Outbreaks. 2013 Dec 12. Edition 1. doi: 10.1371/currents.outbreaks.62df1c7c75ffc96cd59034531e2e836 4. PDF, XML
Authors: Ahmed Abd El Wahed, Department of Virology, University Medical Centre, Goettingen, Germany; Department of Virology, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt. Pranav Patel, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany. Doris Heidenreich, Department of Virology, University Medical Centre, Goettingen, Germany. Frank T. Hufert, Department of Virology, University Medical Centre, Goettingen, Germany. Manfred Weidmann, Department of Virology, University Medical Centre, Goettingen, Germany.
Abstract
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.
Funding Statement
The study was funded by the Federal Ministry of Education and Research (BMBF) (project name: RESCheck, ID: FKN:16SV5436).
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Reverse Transcription Recombinase Polymerase Amplification Assay for the Detection of Middle East Respiratory Syndrome Coronavirus
December 12, 2013 ? Research
Abd El Wahed A, Patel P, Heidenreich D, Hufert FT, Weidmann M. Reverse Transcription Recombinase Polymerase Amplification Assay for the Detection of Middle East Respiratory Syndrome Coronavirus. PLOS Currents Outbreaks. 2013 Dec 12. Edition 1. doi: 10.1371/currents.outbreaks.62df1c7c75ffc96cd59034531e2e836 4. PDF, XML
Authors: Ahmed Abd El Wahed, Department of Virology, University Medical Centre, Goettingen, Germany; Department of Virology, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt. Pranav Patel, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany. Doris Heidenreich, Department of Virology, University Medical Centre, Goettingen, Germany. Frank T. Hufert, Department of Virology, University Medical Centre, Goettingen, Germany. Manfred Weidmann, Department of Virology, University Medical Centre, Goettingen, Germany.
Abstract
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.
Funding Statement
The study was funded by the Federal Ministry of Education and Research (BMBF) (project name: RESCheck, ID: FKN:16SV5436).
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