Real-Time Reverse Transcription-PCR Assay Panel for Middle East Respiratory Syndrome Coronavirus

Lu, Xiaoyan; Whitaker, Brett; Sakthivel, Senthil Kumar K.; Kamili, Shifaq; Rose, Laura; Lowe, Luis; Mohareb, Emad; Elassal, Emad M.; Alsanouri, Tarek; Haddadin, Aktham; Erdman, Dean D.

doi:10.1128/jcm.02533-13

Cited by 159 publications

(198 citation statements)

References 13 publications

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“…Current diagnostic tests for coronavirus include RT‐PCR, real‐time reverse transcription PCR (rRT‐PCR), reverse transcription loop‐mediated isothermal amplification (RT‐LAMP), as well as real‐time RT‐LAMP 51,55–58 …”

Section: Coronavirus and Its Diagnostic Approachesmentioning

confidence: 99%

Recent advances in the detection of respiratory virus infection in humans

Zhang

Wang

Deng

et al. 2020

Journal of Medical Virology

445

363

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Respiratory tract viral infection caused by viruses or bacteria isone of the most common diseases in human worldwide, while those caused by emerging viruses, such as the novel coronavirus, 2019-nCoV that caused the pneumonia outbreak in Wuhan, China most recently, have posed great threats to global public health. Identification of the causative viral pathogens of respiratory tract viral infections is important to select an appropriate treatment, save people's lives, stop the epidemics, and avoid unnecessary use of antibiotics. Conventional diagnostic tests, such as the assays for rapid detection of antiviral antibodies or viral antigens, are widely used in many clinical laboratories. With the development of modern technologies, new diagnostic strategies, including multiplex nucleic acid amplification and microarray-based assays, are emerging. This review summarizes currently available and novel emerging diagnostic methods for the detection of common respiratory viruses, such as influenza virus, human respiratory syncytial virus, coronavirus, human adenovirus, and human rhinovirus. Multiplex assays for simultaneous detection of multiple respiratory viruses are also described. It is anticipated that such data will assist researchers and clinicians to develop appropriate diagnostic strategies for timely and effective detection of respiratory virus infections. K E Y W O R D S adenovirus, coronavirus, diagnostic methods, influenza virus, respiratory syncytial virus, respiratory viral infection, rhinovirus

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Section: Coronavirus and Its Diagnostic Approachesmentioning

confidence: 99%

Recent advances in the detection of respiratory virus infection in humans

Zhang

Wang

Deng

et al. 2020

Journal of Medical Virology

445

363

View full text Add to dashboard Cite

show abstract

“…This detection is performed by targeting regions upstream of the E gene (upE) or within open reading frame (ORF)1b, using upE for screening and ORF1b for confirmation (42). Another real-time RT-PCR assay targeting the MERS-CoV nucleocapsid protein gene was developed and can be used for screening and confirmation (43). When there are discordant results between two real-time RT-PCR assays, the sequencing of an amplicon generated from an appropriate RT-PCR assay should be performed to confirm the test results (44).…”

Section: Diagnosismentioning

confidence: 99%

Middle East Respiratory Syndrome

Alsolamy

2015

Critical Care Medicine

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“…Results submitted for synthetic specimens J and K were not scored, as participants were likely to obtain varying results depending on the gene region and PCR marker used by the testing laboratory. In particular, participants performing MERS‐CoV‐specific testing according to the assay developed by the United States Centers for Disease Control and Prevention (US CDC) would expect a negative result for specimen J, as this assay targets a region of the N gene that is different to the region that was used to design the in vitro RNA transcript included in specimen J . These specimens were included as they could provide interesting information in regard to how participating laboratories handle equivocal test results.…”

Section: Methodsmentioning

confidence: 99%

Proficiency testing for the detection of Middle East respiratory syndrome coronavirus demonstrates global capacity to detect Middle East respiratory syndrome coronavirus

Theis

Lau

Gray

et al. 2018

Journal of Medical Virology

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The first reported case of Middle East respiratory syndrome coronavirus (MERS-CoV) infection was identified in Saudi Arabia in September 2012, since which time there have been over 2000 laboratory-confirmed cases, including 750 deaths in 27 countries. Nucleic acid testing (NAT) is the preferred method for the detection of MERS-CoV. A single round of a Proficiency Testing Program (PTP) was used to assess the capability of laboratories globally to accurately detect the presence of MERS-CoV using NAT. A panel of eleven lyophilized specimens containing different viral loads of MERS-CoV, common coronaviruses, and in vitro RNA transcripts was distributed to laboratories in all six World Health Organization regions. A total of 96 laboratories from 79 countries participating in the PTP, with 76 of 96 (79.2%) reporting correct MERS-CoV results for all nine scored specimens. A further 10 laboratories (10.4%) scored correctly in eight of nine specimens of the PTP. The majority of laboratories demonstrated satisfactory performance in detecting the presence of MERS-CoV using NAT. However, some laboratories require improved assay sensitivity, reduced cross contamination of samples, and improved speciation of coronavirus subtypes for potentially complex clinical specimens. Further PTP and enhanced links with expert laboratories globally may improve the laboratory performance.

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Real-Time Reverse Transcription-PCR Assay Panel for Middle East Respiratory Syndrome Coronavirus

Cited by 159 publications

References 13 publications

Recent advances in the detection of respiratory virus infection in humans

Recent advances in the detection of respiratory virus infection in humans

Middle East Respiratory Syndrome

Proficiency testing for the detection of Middle East respiratory syndrome coronavirus demonstrates global capacity to detect Middle East respiratory syndrome coronavirus

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