key: cord-0811282-rvyeb4c6
authors: Theis, Torsten; Lau, Katherine A.; Gray, Joanna L.; Oxenford, Christopher J.; Walker, Gregory J.; Rawlinson, William D.
title: Proficiency testing for the detection of Middle East respiratory syndrome coronavirus demonstrates global capacity to detect Middle East respiratory syndrome coronavirus
date: 2018-08-27
journal: J Med Virol
DOI: 10.1002/jmv.25266
sha: 289e60f45e35694a7943a6536d25f1b293b28c5b
doc_id: 811282
cord_uid: rvyeb4c6

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.

The WHO identified 133 laboratories from 98 countries in all six WHO regions as potential participants. One hundred and two laboratories confirmed their interest and 99 subsequently confirmed their ability to receive the panel. A total of 96 laboratories located in 79 different countries returned results and participated in the PTP (Table 1 ). The additional verification step was introduced to ensure laboratories had all the documentation and permits in place that were required to import the material into their country.

The proficiency testing (PT) panel consisted of 11 specimens. This was made up of nine specimens (A to I) of inactivated (gamma irradiation; 50 kGy) MERS-CoV, human coronavirus OC43, human coronavirus 229E, and a negative control (specimen I), as well as two synthetic specimens (J and K) containing in vitro RNA transcripts.

RCPAQAP have used similar transcripts in previous PT panels, and have shown that they are safe, stable, and reliable. 8 Following gamma irradiation of the viruses, inactivation was confirmed and viral RNA was quantified using real-time PCR. 8 The relative measure of the concentration of virus-specific target was determined by generating standard curves using a set of MERS-CoV-specific and HCoV-specific primers to quantify the genome equivalents (GE) copies per millilitre of each specimen. 6, 7 Three external referee laboratories confirmed sample characteristics.

Participants were requested to test all specimens of the PT panel and 10-fold MERS-CoV dilution panel. 11, 12 These studies also demonstrated a high overall capacity for laboratories to detect MERS-CoV in China, 11 and worldwide. 12 Similarly, a study performed during the 2015 Korean outbreak reported a 100% MERS-CoV detection score amongst 47 participants. 13 However, the panel in this study only consisted of three specimens, limiting their assessment of assay sensitivity. Both the Korean and Chinese external quality assessments differed from this PTP in that they did not include other human coronaviruses for the assessment of assay specificity. Access to and participation in future quality assurance (QA) programs will provide the opportunity for improvement at a technical level.

Offering an ongoing PTP for the detection of MERS-CoV and other human coronaviruses by PCR will ensure that this capacity will remain appropriate and provide continuing opportunities for improvement. of documentation such as import permits. Obtaining these documents was often time-consuming, and in more than one instance required 8 weeks or longer before specimens could be received. The responses to these challenges need to include early engagement with laboratories, as much local interaction as possible with regional WHO support, and effective communication with customs and courier officials to avoid delays at the dispatch stage of the PTP.

The strong support from the WHO regional offices was key to successfully running the PTP in 96 laboratories from 79 countries. 

Coronavirus diversity, phylogeny and interspecies jumping

Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia

WHO. Middle East respiratory syndrome coronavirus

Laboratory capability for molecular detection and confirmation of novel coronavirus in Europe

The United Kingdom public health response to an

Assays for laboratory confirmation of novel human coronavirus (hCoV-EMC) infections

Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction

Laboratory testing for Middle East respiratory syndrome coronavirus: Interim guidance

Accessed 1

Ebola preparedness: Diagnosis improvement using rapid approaches for proficiency testing

Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus

External quality assessment for the molecular detection of MERS-CoV in China

First international external quality assessment of molecular diagnostics for Mers-CoV

External quality assessment of MERS-CoV molecular diagnostics during the 2015 Korean outbreak

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

We would like to thank all laboratories for participating in this study.The authors are grateful to Julian Druce from the VIDRL in Melbourne, Australia, for the preparation of all virus strains used in this study. We are thankful to Public Health England who provided the MERS-CoV strain that was included in the proficiency testing panel.

The authors declare that there are no conflicts of interest.

Gregory J. Walker http://orcid.org/0000-0003-0044-1394