MERS-CoV and H7N9 Influenza Assay Development on NGDX

Caballero and Armstrong-Spenrath. 59th Medical Wing. 2018

The full report is publicly available and can be downloaded here. The abstract is below:

                                                                           ABSTRACT
"Deadly infectious diseases pose a prevalent danger to war fighters and warrior medics in remote, hostile areas. Infectious agents inevitably hinder the war fighters’ duty performance, even potentially cause mission failures. Therefore a crucial military need is to acquire the capabilities to rapidly detect the threat agents, and to expeditiously devise strategies to counter the threats. The Biomeme two3™ (Biomeme, Inc., Philadelphia, PA) is a light (1.2 lb.), hand-held, fielddeployable real-time polymerase chain reaction (PCR) device that could meet these needs. The device is coupled to an iPhone with unique software for data analysis and transmission to intended recipients. This work reports a comparative research testing and evaluation of this system, focusing on detection of the Middle East Respiratory Syndrome Coronavirus (MERSCoV) and Influenza A virus H7N9 for this assessment. Three Biomeme two3™ instruments were purchased for this work. The reagents specifically designed and set in the appropriate format for Biomeme two3™ and the target templates were also purchased from the manufacturer of the instrument. For MERS-CoV, the detection targets were an orf1a segment and a segment upstream of gene E (termed upE). For H7N9, the target amplicons were in the H7 and N9 genes. The instrument performance was evaluated for template copy numbers that varied from 50 to 500,000 per reaction.

Our results show that Biomeme two3™ can detect the tested targets at various copy numbers, down to 50 copies per reaction. We also tested the MERS-CoV detection reagents for their capacity to amplify the corresponding genomic segments from two nontarget coronaviruses, OC43 and 229E. These tests yielded no amplicons. Likewise, we tested the H7N9 detection reagents to see whether they would amplify the corresponding gene segments from the nontarget influenza A virus H1N1. These PCR reactions also did not produce any amplicons. Together these results show the specificity of the reagents for detection of MERS-CoV and H7N9 by realtime PCR performed on Biomeme two3™.

For comparison we also employed benchtop real-time PCR instruments. Overall, our findings suggest that the Biomeme two3™ system performance in detecting the targets was similar to those of the benchtop instruments."