MERS-CoV and H7N9 Influenza Assay Development on NGDX
Technical Report,01 May 2016,01 Jul 2018
Center for Advanced Molecular Detection JBSA-Lackland United States
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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 capabilitiesto 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, field-deployable 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 detectionreagents 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.
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