268. Rapid Development of a Real-Time RT-PCR Assay for the Detection of the 2012 Novel Coronavirus on the BD MAX™ System
Session: Poster Abstract Session: Diagnostic Microbiology; Novel Molecular Methods
Thursday, October 3, 2013
Room: The Moscone Center: Poster Hall C
Background: In 2012, a novel human coronavirus emerged in the Middle East.  Between April 2012 and March 2013 there have been 17 confirmed cases in Saudi Arabia, Qatar, Jordan, the United Kingdom, and the United Arab Emirates with 11 of these resulting in death.  The genome sequence of the novel coronavirus has revealed that it is a novel betacoronavirus lineage that is most closely related to bat coronaviruses HKU4 and HKU5.  Most of the people infected with this virus had severe acute respiratory illness.  Given the severity of the illness caused by this virus it could pose a significant global health risk if it were to develop efficient human-to-human transfer.

The goal of this study was to demonstrate the ability to rapidly develop a real-time RT-PCR assay on the BD MAX™ System in response to this emerging infectious agent.

Methods: Primers, probes, and a plasmid control were designed for the RNA-dependent RNA polymerase region using three genomes sequences for the novel coronavirus.  For an internal control we used primers and probes that had been developed previously for the human RNaseP gene.  Real-time RT-PCR was performed on the BD MAX™ System with the SuperScript® III Platinum® One-Step qRT-PCR kit.  Analytical sensitivity was determined by serially diluting the plasmid control.  Analytical specificity was determined with 22 common respiratory viruses and bacteria including the four human coronaviruses.  Clinical testing was performed with 20 spiked positive samples and 56 negative clinical samples.

Results: Our assay for the novel coronavirus was able to detect the plasmid control down to 7 copies per reaction in negative clinical sample matrix.  The assay did not cross react with any of the human coronaviruses or other respiratory pathogens tested.  The assay had 100% sensitivity (95% CI 80.0 - 100%) and specificity (95% CI 92.0 - 100%) in the clinical testing performed.

Conclusion: Due in part to the BD MAX™ being an open system we were able to rapidly develop an assay for the novel coronavirus that was highly sensitivity and specific.  Rapid response to emerging infectious agents could be critical in a global outbreak.

Michael Bose, MS1, Amy Sasman, MS1 and Kelly Henrickson, MD1,2, (1)Pediatrics, Medical College of Wisconsin, Milwaukee, WI, (2)Pediatrics, Children's Research Institute, Milwaukee, WI


M. Bose, Becton Dickinson and Company: Research Contractor, Research support

A. Sasman, Becton Dickinson and Company: Research Contractor, Research support

K. Henrickson, Becton Dickinson and Company: Research Contractor, Research support

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